Role of the N+1 Amino Acid Residue on the Deamidation of Asparagine in Pentapeptides

Role of the n+1 amino acid residue on the deamidation of asparagine in pentapeptides

Hasan H. Ince,§F. Aylin SungurKonuklar†, IlkeUgur‡¥Ψ, Maryam Sayadiþ, Ö. AlazOzcan¥, Michael Feig£ and ViktoryaAviyente¥*

§ Department of Chemistry, Michigan State University, East Lansing, Michigan 48824,USA

†Informatics Institute, Computational Science and Engineering Division,Istanbul Technical University, Maslak, Istanbul 34469, Turkey

£ Department of Chemistry and Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan 48824, Turkey

þKimLab,University of Toronto,Toronto, Ontario, M5S 3E1, Canada

‡Université de Lorraine, UMR 7565 SRSMC, Boulevard des Aiguillettes B.P. 70239,F-54506 Vandoeuvre-les-Nancy, France

Ψ CNRS, UMR 7565 SRSMC, Boulevard desAiguillettes B.P. 70239, F-54506 Vandoeuvre-les-Nancy, France

¥Department of Chemistry, Bogaziçi University, Bebek, Istanbul, 34342, Turkey

* Author to whom correspondence should be addressed:

RESULTS

CONFORMATIONAL SAMPLING

Figure S1.Schematic overview of major conformational basins sampled by φ/Ψ backbone torsion angles in nonglycine, nonproline peptide residues.

(Taken from :Feig, M. Is Alanine Dipeptide a Good Model for Representing the Torsional Preferences of Protein Backbones?J. Chem. Theory Comput.2008, 4, 1555-1564.)

Figure S2. Probability histograms for PHI, PSI, PHI2, PSI2

II. RATIONALIZATION OF THE DEAMIDATION MECHANISM

Tetrahedral Intermediate Mechanism

The deamidation mechanisms suggested by Çataket al.1-2 have shown the role of water molecule acting as a catalyst which facilitating the ring closure. For this purpose, the location of the active water molecule was of interest in the simulations. Thus, our analysis focused on the search for pre-reactant structures which might lead to expected transition states and four main parameters have been taken into consideration.

- The C=O---H (H2O) and NH--O (H2O) distances are important to characterize the involvement of water in the mechanisms. These distances are scanned from 3.5 Å to 2.0 Å to locate the pre-reactive complexes.

-The angle between O (H2O), H (H2O) and the corresponding third atom (carbonyl oxygen of asparagine side chain or the amide hydrogen of the n+1 residue) as well as the angle between O (H2O), the HN proton of the backbone and the N itself should display a quasi linear arrangement to facilitate the proton transfer. For this purpose, angles (A) exceeding 120° and 150° have been monitored through the trajectories.

The angles of interest have been calculated as:

cos(A) = ( dXH2 + dHY2 – dXY2 ) / 2*dXH*dHY

A = cos-1[ ( dXH2 + dHY2 – dXY2 ) / 2*dXH*dHY] radians * 57.2957795 degrees*radians-1

There are two possible ways to search for the nearest water as starting from the carbonyl oxygen (OD1) or from the backbone hydrogen (HN). In addition, there are two different types of hydrogen atoms described in TIP3P water model (H1 and H2) in addition to the oxygen (OH2). Thus, the following pathway has been followed:

i)Locate the nearest H1 to OD1, and then find the distance from OH2 of this water to HN.

ii)Locate the nearest H2 to OD1, and then find the distance from OH2 of this water to HN.

iii)Locate the nearest OH2 to HN, and then find the distance from H1 of this water to OD1.

iv)Locate the nearest OH2 to HN, and then find the distance from H2 of this water to OD1.

Two different angles have been monitored with each one of the hydrogen atoms of water (H1 and H2).The above mentioned 4 different pathways (distances) have been combined with 2 different angles yielding a total of 8 alternatives. The snapshots satisfying these requirements have been considered for further analysis (Table S1). As seen in Table S1 the ranking of deamidation rate (GGNGG > GGNMG > GGNIG) is not reflected by the number of structures satisfying the conditions for a pre-reactive complex in the tetrahedral intermediate mechanism (GGNIG > GGNMG > GGNGG).

Table S1. Number of structures for tetrahedral intermediate mechanism (44545 at total)

GGNGG / GGNMG / GGNIG
A1*>120 / A2*>150 / A1*>120 / A2*>150 / A1*>120 / A2*>150
<d4 / <d5 / # / # / # / # / # / #
3.5 / 3.5 / 923 / 135 / 1395 / 235 / 1783 / 331
3.4 / 3.4 / 765 / 126 / 1214 / 224 / 1566 / 304
3.3 / 3.3 / 678 / 117 / 1047 / 216 / 1401 / 286
3.2 / 3.2 / 569 / 104 / 916 / 210 / 1232 / 263
3.1 / 3.1 / 489 / 99 / 796 / 195 / 1091 / 251
3 / 3 / 426 / 92 / 692 / 183 / 959 / 235
2.9 / 2.9 / 358 / 85 / 608 / 175 / 839 / 218
2.8 / 2.8 / 311 / 82 / 546 / 165 / 725 / 209
2.7 / 2.7 / 280 / 79 / 485 / 158 / 638 / 190
2.6 / 2.6 / 235 / 70 / 423 / 152 / 541 / 172
2.5 / 2.5 / 187 / 63 / 358 / 134 / 450 / 156
2.4 / 2.4 / 139 / 53 / 292 / 121 / 368 / 145
2.3 / 2.3 / 109 / 47 / 243 / 106 / 268 / 109
2.2 / 2.2 / 80 / 40 / 169 / 80 / 180 / 77
2.1 / 2.1 / 45 / 30 / 94 / 49 / 105 / 55
2 / 2 / 19 / 14 / 47 / 29 / 37 / 21

A1* is the angle between O (H2O), H (H2O) and the corresponding third atom (carbonyl oxygen of asparagine side chain or the amide hydrogen of the n+1 residue).A2* is the angle between O (H2O), the HN proton of the backbone and the N itself. The above mentioned atoms are expected to display a quasi linear arrangement to facilitate the proton transfer.

Succinimide Intermediate Mechanism

An approach similar to the one in Part A is used for the succinimide-mediated mechanism, except that instead of OD1 the amide nitrogen of asparagine is used. The number of structures satisfying the expected requirements does not match the ranking of deamidation rates (GGNGG > GGNMG > GGNIG).

Table S2. Number of structures for succinimide-mediated mechanism (44545 at total)

GGNGG / GGNMG / GGNIG
A1*>120 / A2*>150 / A1*>120 / A2*>150 / A1*>120 / A2*>150
<d4 / <d6 / # / # / # / # / # / #
3.5 / 3.5 / 225 / 4 / 292 / 11 / 463 / 22
3.4 / 3.4 / 165 / 4 / 225 / 8 / 325 / 9
3.3 / 3.3 / 127 / 3 / 165 / 6 / 239 / 7
3.2 / 3.2 / 84 / 2 / 115 / 3 / 163 / 5
3.1 / 3.1 / 52 / 1 / 84 / 3 / 109 / 5
3 / 3 / 30 / 1 / 55 / 3 / 81 / 4
2.9 / 2.9 / 17 / 0 / 36 / 2 / 53 / 2
2.8 / 2.8 / 12 / 24 / 2 / 34 / 1
2.7 / 2.7 / 9 / 15 / 1 / 20 / 0
2.6 / 2.6 / 6 / 9 / 1 / 12
2.5 / 2.5 / 4 / 4 / 0 / 6
2.4 / 2.4 / 1 / 2 / 3
2.3 / 2.3 / 0 / 2 / 3
2.2 / 2.2 / 0 / 1
2.1 / 2.1 / 0
2 / 2

A1* is the angle between O (H2O), H (H2O) and the corresponding third atom (carbonyl oxygen of asparagine side chain or the amide hydrogen of the n+1 residue).A2* is the angle between the angle between O (H2O), the HN proton of the backbone and the N itself. The above mentioned atoms are expected to display a quasi linear arrangement to facilitate the proton transfer.

Figure S3. Lowest energy closed and extended structures for neutral,anion and NAC structures of GGNGG (B3LYP/6-31++G(d,p)).

Figure S4. Lowest energy closed and extended structures for neutral,anion and NAC structures of GGNMG (B3LYP/6-31++G(d,p)).

Figure S5. Lowest energy closed and extended structures for neutral,anion and NAC structures of GGNIG (B3LYP/6-31++G(d,p)).

Catak, S.; Monard, G.; Aviyente, V.; Ruiz-López, M. F. Computational study on nonenzymatic peptide bond cleavage at asparagine and aspartic acid J. Phys. Chem. A. 2008, 112, 8752–8761.
Catak, S., Monard, G.; Aviyente, V., Ruiz-López, M. F. Deamidation of Asparagine Residues: Direct Hydrolysis versus Succinimide-Mediated Deamidation Mechanisms J. Phys. Chem. A. 2009, 113, 1111-1120.

CARTESIAN COORDINATES.

GGNGG-c

N2.56525703.3947110-0.4667300

H2.88240103.58499700.4880210

H3.11231503.9672430-1.1105750

H2.74274902.3190020-0.6761720

C1.11660103.6990340-0.5749600

H0.74968703.3151510-1.5281270

H0.96388004.7813390-0.5500990

C0.40168303.06269300.6173970

O1.01707102.81635301.6561280

N-0.91915702.81649500.4370010

H-1.35305503.1302490-0.4227240

C-1.78518302.33722501.5022010

H-2.68246902.95840501.5584730

H-1.25081002.43036302.4505190

C-2.25692200.88386601.3757840

O-3.10582000.45186602.1585970

N-1.67899000.13738800.3974270

H-1.07041100.6245820-0.2501680

C-2.1500520-1.18782000.0208630

H-2.9285590-1.45039900.7458830

C-2.7413590-1.2088220-1.4077370

H-3.0092080-2.2410850-1.6422130

H-1.9826750-0.8851030-2.1256190

C-3.9408630-0.2805980-1.5540480

O-3.81334000.8948640-1.9199270

N-5.1459020-0.8130180-1.2427470

H-5.9735350-0.2353960-1.2988690

H-5.2576510-1.7797650-0.9756130

C-1.0642560-2.28109900.1559460

O-1.1836210-3.3573420-0.4348550

N-0.0256210-1.99427200.9793570

H-0.0506280-1.11535601.4797400

C0.9995480-2.96821901.3135800

H0.6333910-3.96154001.0471280

H1.1760060-2.95347502.3932750

C2.3642010-2.78281700.6360960

O3.2539460-3.62332900.8429740

N2.5244220-1.6953110-0.1339910

H1.7790480-1.0121860-0.2075960

C3.7900390-1.2973550-0.7276070

H3.9728900-1.8315830-1.6685580

H4.6118980-1.5496640-0.0516190

C3.82403700.2138950-1.0160550

O4.90015300.6997290-1.4156140

O2.72534100.8481550-0.8243110

Energy (B3LYP/6-31+G(d,p)) = -1324.65525956 Hartree

Zero-point correction= 0.359288 (Hartree/Particle)

Thermal correction to Energy= 0.385710

Thermal correction to Enthalpy= 0.386654

Thermal correction to Gibbs Free Energy= 0.298080

GGNGG-A-c

N3.1917730-3.11192200.5606400

H3.4644560-3.4225200-0.3762080

H3.8114640-3.54845101.2432480

H3.2848360-1.99532900.6070840

C1.7732860-3.47215500.8025860

H1.4330740-2.96046001.7041510

H1.6837010-4.55134300.9545150

C0.9665340-3.0777770-0.4354800

O1.5081550-3.0580250-1.5446370

N-0.3352430-2.7900650-0.2134870

H-0.6959920-2.90724000.7254070

C-1.2998430-2.5966800-1.2851480

H-1.9752720-3.4544860-1.3513850

H-0.7445610-2.5324700-2.2244180

C-2.1854460-1.3535520-1.1537600

O-3.2886750-1.3556130-1.7216010

N-1.7063960-0.3190330-0.4369980

H-0.7475800-0.3348170-0.1080450

C-2.43641700.9285130-0.2438440

H-3.22428000.9569770-0.9992050

C-3.08436500.99542401.1682650

H-3.48320002.00585201.2964900

H-2.31332600.82669501.9246130

C-4.1846090-0.02994601.3908140

O-3.9756250-1.11815701.9456040

N-5.41655900.31874100.9433350

H-6.1764410-0.34411901.0093440

H-5.59090101.19785100.4793400

C-1.56430002.1903490-0.4464420

O-2.22240703.2642190-0.6464990

N-0.25953702.0409480-0.3528520

C0.47416203.2852100-0.5114770

H0.11557604.07227600.1717500

H0.35741403.7149340-1.5198850

C1.97401403.1745190-0.2738320

O2.71132104.1738830-0.4051610

N2.43899601.96772500.0838930

H1.77103401.20565800.1502360

C3.83968801.68969000.3354780

H4.20056502.25253401.2056740

H4.45353702.0074510-0.5149860

C4.10299400.19972100.5877010

O5.2864350-0.15870100.7545200

O3.0721040-0.56431200.6079470

Energy (B3LYP/6-31+G(d,p)) =-1324.16829958Hartree

Zero-point correction= 0.344343 (Hartree/Particle)

Thermal correction to Energy= 0.370698

Thermal correction to Enthalpy= 0.371642

Thermal correction to Gibbs Free Energy= 0.283368

GGNGG-o

N6.15378400.1470700-0.3973660

H6.38238900.01696800.5934660

H5.40581600.8826320-0.4527350

H6.99210100.4619300-0.8899920

C5.6262890-1.1323360-0.9590770

H6.4228080-1.8792110-0.9390730

H5.3211810-0.9563820-1.9901780

C4.4763460-1.5667640-0.0509830

O4.5444800-1.35644201.1629100

N3.4120230-2.1408130-0.6505540

H3.4222980-2.2985120-1.6498990

C2.2444460-2.54254500.1192280

H2.5483890-3.09984801.0073210

H1.6310060-3.1990330-0.5021420

C1.3696660-1.37740700.6064250

O0.6210110-1.52713601.5732410

N1.4402980-0.2200180-0.1052170

H2.1707270-0.1353000-0.8033140

C0.83358901.01354600.3821210

H0.90429401.02855601.4741570

C1.55531302.2528910-0.2050440

H1.07675803.14281500.2076600

H1.42849702.2704780-1.2917290

C3.05143202.25074500.0663890

O3.80740601.5390390-0.6296400

N3.50044603.02068301.0657650

H2.87974903.59487001.6178650

H4.48725103.04123401.2875590

C-0.67302501.14795100.0723990

O-1.34796601.97777900.6925100

N-1.16223000.3744820-0.9168470

H-0.5565230-0.3295670-1.3157820

C-2.55503000.4105690-1.3134740

H-2.62597200.0403660-2.3404950

H-2.90142001.4467040-1.3046260

C-3.4602000-0.4672570-0.4289690

O-3.0213070-1.46779200.1575970

N-4.7471540-0.0868880-0.3930400

H-5.07479300.7521050-0.8648590

C-5.8012650-0.81423700.2948500

H-5.5813810-0.89241701.3656510

H-5.8815990-1.8366850-0.0917900

C-7.1622590-0.09769900.1130400

O-8.1446380-0.65369600.6739000

O-7.16693000.9659910-0.5702280

Energy (B3LYP/6-31+G(d,p)) = -1324.64328256Hartree

Zero-point correction= 0.361387 (Hartree/Particle)

Thermal correction to Energy= 0.387819

Thermal correction to Enthalpy= 0.388764

Thermal correction to Gibbs Free Energy= 0.299938

GGNGG-A-o

N3.2136550-2.4450380-1.4513410

H3.4940440-1.9885080-2.3220820

H2.2572090-1.9987590-1.1621500

H3.0553980-3.4286140-1.6810810

C4.2848720-2.3342260-0.4007460

H5.2581390-2.4239750-0.8862460

H4.1523580-3.15771500.2975460

C4.1501980-1.03532100.4010800

O3.7243740-1.05862901.5570910

N4.46978000.1199630-0.2340780

H4.91851600.0991960-1.1401010

C4.23524101.40804000.4106380

H4.63879102.1904080-0.2361060

H4.74764301.45666601.3737670

C2.74309601.67903700.6865530

O2.40189902.31522001.6900970

N1.89187601.1959910-0.2446320

H2.29331200.6267360-0.9818430

C0.48203400.9019760-0.0153130

H0.23239201.21225100.9996260

C-0.45166101.6173180-1.0352080

H-1.38325601.0503880-1.1007320

H0.02203601.6119740-2.0210340

C-0.76206003.0584580-0.6572540

O0.05201003.9798570-0.8342320

N-1.97687503.2598180-0.0994450

H-2.22645004.20010900.1767020

H-2.67447902.52056900.0026760

C0.2707360-0.6253680-0.1389620

O0.9695030-1.2232100-1.0542080

N-0.6476490-1.10867400.6425730

C-1.0911600-2.47537300.4394640

H-0.8219380-3.10811401.2969340

H-0.6593900-2.9499470-0.4531320

C-2.6185320-2.56481800.2971970

O-3.1875210-3.66498700.1455880

N-3.2538400-1.38616500.3353820

H-2.6765320-0.55934800.4733400

C-4.6721500-1.15570600.1590480

H-5.2519310-1.58746500.9846980

H-5.0317580-1.6302210-0.7613410

C-4.97712200.36218000.0888730

O-3.99816401.15813800.2155780

O-6.18506100.6681030-0.0837720

Energy (B3LYP/6-31+G(d,p)) = -1324.15673581Hartree

Zero-point correction= 0.346228 (Hartree/Particle)

Thermal correction to Energy= 0.372037

Thermal correction to Enthalpy= 0.372981

Thermal correction to Gibbs Free Energy= 0.285921

NAC-GGNGG-A-o

N3.7747650-1.25017601.7043440

H2.8863140-1.04180302.1658010

H3.9627230-0.35183201.0700240

H4.4946880-1.31798002.4261040

C3.6938210-2.53238600.9416630

H3.4084130-3.32993001.6301510

H4.6852380-2.74320400.5426490

C2.7296870-2.4336250-0.2500810

O3.1416240-2.0952990-1.3615710

N1.4413200-2.76892800.0084190

H1.2085810-3.12855000.9251160

C0.4340060-2.8778680-1.0326480

H0.1484870-3.9240590-1.1735720

H0.8866430-2.5254910-1.9635950

C-0.8711190-2.1078200-0.8097350

O-1.9158150-2.5931050-1.2899450

N-0.8453400-0.9395110-0.1474140

H0.0463990-0.53641700.1487080

C-2.0856760-0.1485650-0.0072500

H-2.5692440-0.1642200-0.9853830

C-3.0361910-0.76632201.0585910

H-3.0335790-0.13914201.9508280

H-2.6674960-1.75729101.3414460

C-4.4748020-0.92570200.5807010

O-5.4283490-0.35590001.1356890

N-4.6356380-1.7520350-0.4781770

H-5.5560800-1.8803970-0.8744540

H-3.8211440-2.1640570-0.9306330

C-1.72504301.32775500.3049560

O-1.51854901.65073501.5160430

N-1.65781102.0908490-0.7770470

C-1.24362603.4630430-0.5251350

H-1.64326803.87559200.4115270

H-1.59991704.1053160-1.3420510

C0.27707403.6713230-0.4459180

O0.77017204.7265050-0.0083560

N1.00796602.6373140-0.9146220

H0.45674001.8114150-1.1292270

C2.42412002.4547780-0.6860690

H2.80545003.3149460-0.1290570

H2.97992702.4084880-1.6303930

C2.71990101.16994200.1050470

O3.95994500.92721800.3181600

O1.77489500.43968500.4865760

Energy (B3LYP/6-31+G(d,p)) = -1324.1570968 Hartree

Zero-point correction= 0.346772 (Hartree/Particle)

Thermal correction to Energy= 0.371920

Thermal correction to Enthalpy= 0.372864

Thermal correction to Gibbs Free Energy= 0.290050

GGNMG-c

N-2.78627701.88336502.5928960

C-1.47068902.20095803.2065540

C-0.44504001.17499702.7275590

O-0.80352000.14937602.1369150

H-1.55803902.14682104.2945400

H-1.18825403.21843302.9299620

H-2.85229302.20933401.5598360

H-3.54865802.31233303.1196230

H-2.92284000.86736902.5921420

N0.84757201.45532102.9973700

C1.89824200.47197302.7629680

C2.19439500.17154501.2884990

O2.7353780-0.89244700.9762080

H1.07807902.29601903.5115900

H1.6380660-0.48294503.2266600

H2.81466100.83783803.2312730

N1.82479401.12151700.3957590

C1.98991801.0264310-1.0527460

C1.0666190-0.0118840-1.7276580

O1.2555840-0.3163620-2.9101740

C3.44922800.8564050-1.4940270

C4.26057402.1251730-1.2569980

N5.57686301.9385820-1.0089550

O3.75363803.2528820-1.3184070

H1.43012801.97779600.7620310

H1.65128401.9911950-1.4448610

H3.45361100.6611230-2.5715080

H3.9036880-0.0048860-1.0008290

H6.18398202.7413930-0.9158060

H5.99121601.0191550-0.9699660

N0.0219370-0.4793380-1.0000130

C-0.9553700-1.4132740-1.5494330

C-2.2712070-0.7422870-2.0051410

O-3.0997920-1.4041610-2.6502730

C-1.2713730-2.5433670-0.5465330

C-0.0579790-3.4334200-0.2666130

S-0.5182410-4.77030900.9108870

C1.1023470-5.59619401.0989650

H-0.0474340-0.2342880-0.0166600

H-0.5127460-1.8409960-2.4529100

H-2.0846710-3.1398020-0.9694090

H-1.6399490-2.10400900.3885830

H0.7604980-2.85490200.1714900

H0.2994340-3.8935730-1.1939390

H1.4526870-5.99012400.1415470

H0.9584690-6.42560901.7947940

H1.8440670-4.90782901.5117340

N-2.45213800.5399960-1.6496450

C-3.69261401.2771550-1.8185380

C-3.85841402.3363670-0.7085090

O-4.95001802.9358240-0.6531780

O-2.84604802.51675600.0586420

H-1.77025400.9988590-1.0565050

H-4.53955500.5866410-1.7865510

H-3.72617601.7850790-2.7909170

Energy (B3LYP/6-31+G(d,p)) = -1840.79251236Hartree

Zero-point correction= 0.445606 (Hartree/Particle)

Thermal correction to Energy= 0.478096

Thermal correction to Enthalpy= 0.479040

Thermal correction to Gibbs Free Energy= 0.376008

GGNMG-A-c

N-4.88078000.08766201.3543920

C-4.29006801.43292301.5671480

C-2.85548801.24489402.0642840

O-2.53991700.21613402.6668670

H-4.86729501.97455702.3215480

H-4.33508401.98476900.6270030

H-4.3765320-0.41534600.4938100

H-5.88611000.14575901.1890450

H-4.7198040-0.48823802.1850140

N-2.00592902.26616701.8023240

C-0.62436502.27761902.2595020

C0.45327002.08461801.1781780

O1.63927602.07232901.5391740

H-2.37972303.11030001.3861540

H-0.50059001.47810802.9918760

H-0.41058903.22712702.7585530

N0.02376201.9896390-0.0926090

C0.84249601.7790870-1.2916230

C1.32554200.3154220-1.5266250

O2.14818600.1787390-2.4924390

C1.98958502.7943600-1.4001510

C1.49562004.2154720-1.6214150

N2.12976005.1798830-0.9119960

O0.58812304.4860270-2.4216880

H-0.97593801.9071410-0.2150890

H0.16691401.9903090-2.1279500

H2.57452102.5094330-2.2801580

H2.64163202.7349210-0.5275260

H1.88427806.1497470-1.0561100

H2.86700604.9641950-0.2579490

N0.7783950-0.6073630-0.7628370

C1.1409500-1.9843670-1.0566740

C-0.0772230-2.8291680-1.4803090

O0.0555790-4.0019160-1.8852830

C1.7979430-2.66877400.1683450

C3.1673310-2.07148300.4905610

S3.8935720-2.87802201.9784290

C5.4518870-1.93230402.1234020

H1.8429270-2.0521240-1.9009290

H1.8995320-3.7401930-0.0354000

H1.1307170-2.55429201.0325030

H3.0806890-0.99926400.6875010

H3.8559090-2.2162560-0.3492050

H6.0716580-2.06388101.2326210

H5.9876310-2.32349002.9911500

H5.2482730-0.86963802.2786700

N-1.2696110-2.2206590-1.3714660

C-2.5422120-2.8565970-1.6483740

C-3.7228510-2.1081840-1.0098220

O-4.8370770-2.6713270-1.0227700

O-3.4681940-0.9498820-0.5202820

H-1.2622330-1.2707080-1.0083870

H-2.5365340-3.8847140-1.2739290

H-2.7298960-2.9191140-2.7295260

Energy (B3LYP/6-31+G(d,p)) = -1840.30280836Hartree

Zero-point correction= 0.430622 (Hartree/Particle)

Thermal correction to Energy= 0.462905

Thermal correction to Enthalpy= 0.463850

Thermal correction to Gibbs Free Energy= 0.360410

GGNMG-o

N6.37575103.78486900.9769540

H6.56911803.15852001.7792990

H5.78315204.54566801.3187980

H7.24843604.19537000.6384920

C5.68587902.9539400-0.0583470

H6.43500302.5921120-0.7648020

H4.95666903.5645950-0.5895240

C5.05401201.76708600.6825520

O5.42696501.50848001.8361540

N4.14599501.06457900.0016850

H3.86437101.3234720-0.9402720

C3.5016670-0.11860000.5400320

H2.97920900.11620701.4734520

H4.2418630-0.89558300.7641290

C2.5091270-0.6389220-0.4998410

O2.3994610-0.1031910-1.6074410

N1.7587710-1.6967660-0.1091130

H2.0422280-2.22419200.7159060

C0.9230520-2.4154530-1.0689730

H1.4887970-2.5809930-1.9917700

C0.4740230-3.7773100-0.4862480

H-0.1073330-4.2936980-1.2524780

H-0.1790140-3.60839500.3764900

C1.6417590-4.62391000.0051480

O2.3165600-4.25983100.9825900

N1.8935680-5.7653280-0.6641120

H2.6573090-6.3581800-0.3674490

H1.3446970-6.0564630-1.4592660

C-0.3366530-1.6352040-1.5006380

O-0.8740150-1.8997760-2.5839390

N-0.8120580-0.7322710-0.6237860

H-0.2772650-0.50608700.2090210

C-1.95781000.1229070-0.8937950

H-1.95372600.3679360-1.9610610

C-3.3080100-0.5750650-0.5855090

H-3.3529720-1.4753960-1.2069640

H-4.12057800.0819690-0.9141140

C-3.4961240-0.94432500.8873290

H-3.5342540-0.04991401.5171400

H-2.6747000-1.57827901.2350440

S-5.0720000-1.86850601.1039170

C-5.0055070-2.18193502.9038760

H-5.9100180-2.73534503.1651380

H-4.9877330-1.24268903.4624190

H-4.1308490-2.78458803.1613440

C-1.76366501.4142130-0.0741710

O-0.97810101.44092200.8900100

N-2.50459402.4613100-0.4565750

H-3.10795002.4296170-1.2754020

C-2.49948103.75952700.1999290

H-1.48485504.17193000.2287730

H-2.84223903.67092701.2370280

C-3.42522104.7448180-0.5566700

O-3.49074105.9058830-0.0722290

O-4.02259004.3002910-1.5783280

Energy (B3LYP/6-31+G(d,p)) = -1840.77876397Hartree

Zero-point correction= 0.445297 (Hartree/Particle)

Thermal correction to Energy= 0.478713

Thermal correction to Enthalpy= 0.479658

Thermal correction to Gibbs Free Energy= 0.371290

GGNMG-A-o

N-8.1692130-0.43389800.4297230

H-8.6326700-0.68126101.3063480

H-7.6746090-1.26266400.0481270

H-8.8915890-0.1674040-0.2433230

C-7.13188800.63037800.5987830

H-6.84590500.67572601.6508820

H-7.54281701.59362800.2987320

C-5.92462500.2061110-0.2499820

O-5.8901510-0.9400360-0.7162400

N-4.97202801.1324400-0.4215020

H-5.06559802.02025400.0547250

C-3.70874400.8376890-1.0675970

H-3.29967901.7705580-1.4634730

H-3.88974300.1669270-1.9111000

C-2.68738200.1876520-0.1081660

O-2.9662480-0.04512901.0735010

N-1.5062690-0.0980610-0.6896250

H-1.29889500.2536830-1.6223480

C-0.3096050-0.56627800.0075570

H-0.4390460-0.39398601.0763840

C-0.0580800-2.0753290-0.2445050

H0.9079130-2.34117400.1940870

H-0.0019360-2.2501540-1.3238210

C-1.1015550-2.97100000.4041690

O-1.1068400-3.18647901.6248750

N-2.0220000-3.5215090-0.4233530

H-2.7507290-4.1079120-0.0400050

H-2.0303310-3.3352030-1.4146600

C0.89637200.2559460-0.5228830

O0.82311100.6318680-1.7416520

N1.85480900.43786900.3605420

C3.03107701.1679040-0.0862540

H2.84662501.6977040-1.0315910

C3.45604602.20030500.9852960

H4.42519402.63183200.7121380

H3.59302201.66731001.9354360

C2.43642903.31470801.2280710

H2.71465803.88822902.1180820

H1.44101602.88819701.3849300

S2.35343504.4937970-0.1886600

C1.11793805.67291300.4648820

H1.47287206.13752101.3887400

H0.16173805.17506500.6460590

H0.97835006.4472190-0.2926680

C4.24024900.2565980-0.3800640

O5.28872600.7186530-0.8776720

N4.0896410-1.0365240-0.0606690

H3.1994980-1.33064100.3322050

C5.0899500-2.0675200-0.2562500

H5.9973330-1.84474900.3184710

H5.3915510-2.1190840-1.3095810

C4.5596650-3.45665100.1749900

O3.3742240-3.52039200.6068290

O5.3795720-4.40887800.0493190

Energy (B3LYP/6-31+G(d,p)) = -1840.28753786 Hartree

Zero-point correction= 0.431751 (Hartree/Particle)

Thermal correction to Energy= 0.464842

Thermal correction to Enthalpy= 0.465786

Thermal correction to Gibbs Free Energy= 0.358054

NAC-GGNMG-A-o

N2.68954901.88588000.0595250

H2.32319202.8599130-0.1297800

H2.74463501.75850801.0730460

H1.95758101.2150480-0.3167450

C4.02219901.5908880-0.5436160

H4.75128602.3089810-0.1642790

H3.94146801.6930660-1.6258610

C4.39739000.1768870-0.0889110

O4.3997440-0.09014501.1164290

N4.6103520-0.7454450-1.0573900

H4.5874710-0.4537020-2.0258750

C4.6309660-2.1732860-0.7634550

H5.2616150-2.36397600.1072440

H5.0635050-2.6998260-1.6165530

C3.2425940-2.7828960-0.4525270

O3.1768250-3.9397570-0.0081320

N2.2029350-1.9742500-0.6919970

H2.3749680-1.0540980-1.0904110

C0.7932150-2.1036980-0.3371980

H0.3196990-2.8902780-0.9338160

C0.6076930-2.45326801.1555130

H0.7727500-1.56880201.7778660

H1.3633340-3.19813101.4287520

C-0.7355460-3.10195101.4545750

O-1.1869280-4.02300700.7604600

N-1.3656570-2.67316202.5737650

H-2.2516110-3.08539402.8320340

H-1.0378820-1.87766203.1002610

C0.1479970-0.7270690-0.6974770

O0.95544200.1955850-1.1192070

N-1.1420290-0.6454290-0.5457640

C-1.73644900.6393700-0.9251090

H-1.19712901.0604500-1.7851000

C-3.20005200.4512980-1.3893560

H-3.1607900-0.1453840-2.3095090

H-3.60749201.4324540-1.6665370

C-4.1349030-0.2332650-0.3905990

H-4.24955100.36967600.5118880

H-3.7390510-1.2127350-0.1113260

S-5.7964030-0.4708580-1.1544670

C-6.6724790-1.27548000.2342880

H-6.6968510-0.62136901.1097730

H-6.1984100-2.22510600.4958580

H-7.6958330-1.4670180-0.0960540

C-1.63123601.67607700.2125760

O-2.23181201.55434901.2948200

N-0.84967002.7489060-0.0515730

H-0.27341002.7530890-0.8840990

C-0.61438303.82519400.8973500

H-0.44384003.40660601.8968520

H-1.49193104.47655500.9756790

C0.60817204.69177900.5239770

O0.71875405.77838901.1349420

O1.41238704.2350710-0.3578130

Energy (B3LYP/6-31+G(d,p)) = -1840.3006171 Hartree

Zero-point correction= 0.432074 (Hartree/Particle)

Thermal correction to Energy= 0.463896

Thermal correction to Enthalpy= 0.464841

Thermal correction to Gibbs Free Energy= 0.364294

GGNIG-c

N-5.56354400.7469170-0.3353780

H-5.54768600.4688430-1.3235960

H-4.86363701.5792570-0.1791100

H-6.50705801.0605350-0.1053060

C-5.1630490-0.43058200.4846950

H-6.0424170-1.04905700.6797700

H-4.7612220-0.07453601.4335480

C-4.1545180-1.2505070-0.3254330

O-4.3105410-1.3715090-1.5459740

N-3.1792590-1.84150500.3886050

H-3.0309400-1.54144201.3468510

C-2.0107660-2.4388610-0.2308320

H-1.9508150-2.1214350-1.2744900

H-2.0650880-3.5352430-0.2177140

C-0.7670510-2.00448500.5494970

O-0.8585220-1.50363101.6747850

N0.4253670-2.2066710-0.0661140

H0.4677160-2.8273530-0.8736290

C1.6676850-2.06476200.6915440

H1.5665940-2.56798501.6583470

C2.8597300-2.6722190-0.0864240

H3.7513180-2.57448100.5363310

H3.0307280-2.1023470-1.0061150

C2.6255260-4.1186740-0.5077360

O1.7103330-4.4075650-1.2961820

N3.4593550-5.04456900.0036160

H3.3509670-6.0134510-0.2653390

H4.2078240-4.80576600.6369840

C2.0087830-0.59770801.0388950

O2.5721380-0.33654802.1054830

N1.71155600.32826500.0935480

H1.3239110-0.0122070-0.7784040

C2.12898301.72885000.1907720

H2.80906301.77531701.0427920

C2.90221302.1516070-1.0994580

H3.56841601.3033840-1.3094630

C1.97274502.3396480-2.3103520

H2.56683702.4954840-3.2163380

H1.32491603.2141520-2.1825390

H1.32901001.4716720-2.4922280

C3.79501403.3950530-0.9036410

H3.16976804.2521000-0.6312190

H4.23648503.6265270-1.8815220

C4.92758303.23107000.1175660

H5.57107704.11738700.1204190

H5.55628702.3648220-0.1211830

H4.55239203.10140501.1382910

C0.97226802.69313800.5278590

O1.22203103.84302300.9260980

N-0.27445102.23268800.3434500

H-0.43953701.28595200.0190690

C-1.49070803.00507900.5206140

H-1.44633203.9416700-0.0468560

H-1.64668803.27227001.5721660

C-2.68353802.16649600.0292240

O-3.84402802.67558700.1968700

O-2.45146901.0493670-0.4853760

Energy (B3LYP/6-31+G(d,p)) = -1481.91866538 Hartree

Zero-point correction= 0.472675 (Hartree/Particle)

Thermal correction to Energy= 0.504203

Thermal correction to Enthalpy= 0.505147

Thermal correction to Gibbs Free Energy= 0.407675

GGNIG-A-c

N-5.22681501.79284000.0037520

H-5.21408502.0223450-0.9956800

H-4.39661602.35232200.4747850

H-6.11228902.10889600.4001980

C-5.06203200.32278500.1488580

H-6.0115250-0.1705190-0.0749810

H-4.77052800.10435701.1756390

C-3.9948990-0.1281920-0.8514810

O-3.77181000.5542300-1.8604620

N-3.3865660-1.2932750-0.5708710

H-3.5682240-1.74987500.3159190

C-2.1918640-1.7465050-1.2721120

H-1.7961630-0.9167060-1.8588670

H-2.4270940-2.5686570-1.9595030

C-1.1910010-2.2666950-0.2346620

O-1.5889580-3.01136600.6765570

N0.0852140-1.8817970-0.4075710

H0.2866310-1.0721300-0.9937090

C1.1592490-2.13943300.5471780

H0.7044770-2.42446501.5004310

C2.1039920-3.27322200.0717440

H2.9492920-3.30212700.7652600

H2.4808030-3.0308640-0.9255200

C1.4438510-4.6408310-0.0052020

O0.9989770-5.1045030-1.0648750

N1.3709210-5.32927201.1611080

H0.8994590-6.22280601.1840010

H1.7279980-4.95701802.0284300

C1.9925540-0.84575800.7868170

O2.8412990-0.91221801.7330920

N1.71674300.1442860-0.0367530

C2.46319101.38610800.1290650

H3.20607801.29049500.9313500

C3.22207201.7267840-1.1971060

H3.69669000.7761340-1.4802330

C2.25981502.1199180-2.3293930

H2.79399702.1851240-3.2842760

H1.80308703.0995400-2.1406000

H1.45635901.3857130-2.4388390

C4.34751502.7721180-1.0543990

H3.92099203.7175460-0.7022120

H4.74522702.9623750-2.0611130

C5.51265302.3595220-0.1447170

H6.31093503.1098670-0.1727710

H5.94503201.4028750-0.4633180

H5.20278402.25162800.9001380

C1.54182102.54248600.5710580

O1.98939403.66820000.8711680

N0.23217502.24575200.6117530

H-0.02262101.28324000.4063750

C-0.82586003.16232700.9833260

H-0.88424704.00505400.2858560

H-0.64544403.58828101.9797750

C-2.17557702.42698301.0035620

O-3.21885703.17149200.9648150

O-2.18199201.17782201.0494950

Energy (B3LYP/6-31+G(d,p)) = -1481.43031029Hartree

Zero-point correction= 0.457185 (Hartree/Particle)

Thermal correction to Energy= 0.488990

Thermal correction to Enthalpy= 0.489934

Thermal correction to Gibbs Free Energy= 0.389917

GGNIG-o

N6.5836440-3.54593600.1335230

H5.7026760-3.7026530-0.3921620

H6.6296670-4.18928300.9265650

H7.3627290-3.7641180-0.4923860

C6.5739330-2.10872500.5488050

H7.5684220-1.68343600.4187610

H6.2953850-2.04986101.6020050

C5.5108530-1.4096710-0.3100340

O4.7371990-2.0962320-0.9890680

N5.4946860-0.0700170-0.2546830

H6.12619800.40195000.3796550

C4.46780700.7173490-0.9048790

H4.21704100.2518950-1.8615210

H4.87036401.7122520-1.1107250

C3.19494600.8501230-0.0460520

O3.12718100.40010701.0982600

N2.19093601.5281290-0.6573140

H2.28332801.7466750-1.6422280

C0.87521801.7197590-0.0630140

H0.97391601.48991800.9986910

C0.40920503.1784770-0.2244700

H-0.61781303.26976500.1482390

H0.39288703.4537040-1.2833280

C1.26528404.14191000.5944400

O1.58188303.89233301.7633690

N1.62449005.2868980-0.0274030

H2.14494305.98741100.4831190

H1.36356005.4868640-0.9813450

C-0.14643400.7775830-0.7311360

O-0.12439300.5919090-1.9537420

N-1.04105200.21367600.1092120

H-1.06301500.51920801.0760150

C-2.2127150-0.5379100-0.3255610

H-2.3229090-0.3663920-1.4002470

C-2.0752960-2.0766490-0.0957630

H-3.0260730-2.5104720-0.4358870

C-0.9553570-2.6508120-0.9765100

H-0.9217100-3.7413600-0.9027890

H0.0228200-2.2633280-0.6712560

H-1.1085670-2.3919920-2.0289730

C-1.9000500-2.43706501.3939270

H-2.6633800-1.91619501.9840400

H-0.9290590-2.06115501.7434550

C-2.0000910-3.93938301.6919100

H-1.9419810-4.12021002.7705070

H-1.1937990-4.50996101.2203440

H-2.9532010-4.34929301.3367930

C-3.43821100.04408600.4088290

O-3.30537600.69731701.4588570

N-4.6272480-0.2285610-0.1460340

H-4.7111080-0.7439230-1.0189350

C-5.90996500.17223500.4098920

H-5.96790701.26268500.5025110

H-6.0432960-0.24387501.4148260

C-7.0652230-0.3184950-0.4979850

O-8.2254250-0.0257600-0.1030120

O-6.7450370-0.9598010-1.5395670

Energy (B3LYP/6-31+G(d,p)) = -1481.90349759 Hartree

Zero-point correction= 0.472594 (Hartree/Particle)

Thermal correction to Energy= 0.505741

Thermal correction to Enthalpy= 0.506686

Thermal correction to Gibbs Free Energy= 0.400069

GGNIG-A-o

N3.1238430-4.2016780-1.5203580

C3.6806330-2.8887530-1.9727540

C3.8969620-2.0409070-0.7124230

O3.8943550-2.59256800.3934620

H2.9953590-2.4302990-2.6846300

H4.6387720-3.0703220-2.4630930

H2.1008240-4.2166170-1.5493600

H3.4075260-4.3247860-0.5324690

H3.4648530-4.9889280-2.0762100

N4.1006540-0.7269070-0.9143060

C4.43789300.19502200.1622980

C3.27687001.00375700.7616010

O3.52620301.76962501.7052600

H4.1090420-0.3759150-1.8640290

H5.17634700.9102280-0.2079020

H4.8961120-0.36913400.9768280

N2.06698700.83621400.2087610

C0.82499001.42590800.6937660

C-0.29049800.35742800.5346080

O-0.1383900-0.4712720-0.4314390

C0.42519202.6919470-0.1132850

C1.43100003.8302710-0.0344970

N1.28985904.67715901.0153820

O2.32270403.9831970-0.8809910

H1.91469800.1122860-0.4892190

H0.94817101.68521801.7465990

H-0.54895403.02233600.2595090

H0.31765002.4217070-1.1671000

H1.96480505.41690801.1513960

H0.57979004.54632201.7202680

N-1.27122700.46098301.4005810

C-2.3646160-0.50701801.2419620

C-3.38301100.10923500.2610700

O-4.33436400.82722500.6264790

C-3.0122170-0.86383902.5972640

C-1.9732750-1.34157903.6374680

C-4.1480580-1.88740302.4198220

C-1.2229830-2.63895903.3003820

H-1.9917520-1.43287100.7797770

H-3.45407500.05957102.9941690

H-2.4951800-1.47348804.5956020

H-1.2428130-0.53950803.7872810

H-4.9496460-1.48484201.7937630

H-4.5834930-2.15487403.3899490

H-3.7906280-2.81235201.9517730

H-0.6472800-2.54437402.3733570

H-1.9024960-3.49059403.1855130

H-0.5171430-2.89031004.1003190

N-3.1395730-0.1536860-1.0415450

C-3.88817200.4429740-2.1338000

C-3.35499900.0009010-3.5170000

O-4.00422400.4435420-4.5067840

O-2.3336230-0.7414160-3.5470280

H-2.2653720-0.6095200-1.2936030

H-3.84757901.5395090-2.0845950

H-4.94691100.1694780-2.0640520

Energy (B3LYP/6-31+G(d,p)) = -1481.41093131Hartree

Zero-point correction= 0.458415 (Hartree/Particle)

Thermal correction to Energy= 0.491153

Thermal correction to Enthalpy= 0.492097

Thermal correction to Gibbs Free Energy= 0.387564

NAC-GGNIG-A-o

N-5.18246603.9838610-0.2603180

H-6.01301004.51440200.0119090

H-5.43644303.3761080-1.0639870

H-4.46681004.6517400-0.5546630

C-4.70726703.06512300.8215280

H-5.23323103.29353001.7484300

H-3.63833103.21646400.9730990

C-4.98787701.63321000.3411490

O-5.44961401.4551320-0.7919630

N-4.68320500.65404201.2098320

H-4.35580000.91005502.1334820

C-4.9361230-0.75593300.9419900

H-5.6866650-0.82828200.1521620

H-5.3498110-1.21750201.8416870

C-3.7283600-1.59991800.5089680

O-3.8945270-2.82060600.3523040

N-2.5654200-0.96225900.3245650

H-2.47849300.04065500.4714560

C-1.3121630-1.5930000-0.0791020

H-1.2220590-2.55856500.4235800

C-1.3089690-1.8066920-1.6244440

H-1.1300950-0.8410250-2.1092720

H-2.2970910-2.1644300-1.9206850

C-0.3158010-2.8507500-2.1225510

O-0.6829810-4.0207000-2.3374390

N0.9474370-2.4372030-2.3602710

H1.3240520-1.5736380-1.9518320

H1.6184840-3.1553730-2.6024040

C-0.1742140-0.63967000.3556020

O-0.45426400.61053800.3312820

N0.9592410-1.22663800.6560860

C2.0564900-0.31812800.9349760

H1.73463000.48493201.6191780

C3.2446840-1.04146301.6413060

H4.0254470-0.27975301.7875920

C2.8082130-1.52442203.0336880

H2.0325090-2.29278702.9510470

H2.3999580-0.69587103.6246140

H3.6489510-1.94572503.5940440

C3.8642000-2.17421900.7969840

H4.0159020-1.8144470-0.2279000

H3.1485390-3.00226800.7273080

C5.2053650-2.69479001.3335970

H5.9392510-1.88350401.4174060

H5.6249890-3.45235800.6620240

H5.1055300-3.15503602.3223790

C2.54172500.4040040-0.3436670

O2.3727360-0.0533920-1.4932740

N3.19473401.5628450-0.1402110

H3.29308101.95688800.7905530

C3.76293002.3922710-1.1885140

H2.99523702.6726470-1.9195290

H4.54052201.8490220-1.7385650

C4.37989003.6823930-0.5950150

O4.91121604.4629800-1.4314170

O4.29895403.84148500.6567760

Energy (B3LYP/6-31+G(d,p)) = -1481.4142274 Hartree

Zero-point correction= 0.459301 (Hartree/Particle)

Thermal correction to Energy= 0.491381

Thermal correction to Enthalpy= 0.492325

Thermal correction to Gibbs Free Energy= 0.389989