2
Supplemental Table 1 Domains and conserved regions or amino acids of AtHXKs with respect to ScHXK2
YHK2 HXK1 HXK2 HXK3 HKL1 HKL2 HKL3
Large domain 13-76 27-91 27-91 32-94 27-91 27-91 28-92
212-457 230-475 230-475 235-476 230-483 231-479 231-470
Small domain 77-211 92-229 92-229 95-234 92-229 92-230 93-230
458-486 476-496 476-502 477-493 484-498 480-502 471-493
Phosphate 1 82-103 97-118 cons cons not cons not cons not cons
Loop 1 87-91 102-106 cons cons uncer uncer not cons
Loop 2 115-124 not cons not cons not cons not cons not cons not cons
Sugar binding core 148-167 167-186 cons cons uncer uncer not cons
Loop 3 158-163 177-182 cons cons cons cons not cons
Loop 4 174-178 193-197 cons cons cons cons not cons
Connect 1 203-223 222-242 cons cons not cons not cons not cons
Phosphate 2 229-248 248-267 cons cons cons cons not cons
Thr234 Thr253 cons cons cons cons Met
Adenosine 411-439 425-461 cons cons insert insert not cons
Ser419 Gly441 Gly Ala Gly Gly Gly
Connect 2 453-473 471-491 cons cons cons cons cons
Hydrophobic Ile85 Leu100 Leu100 Leu103 Leu100 Leu100 Val101
Channel Leu87 Leu102 Leu102 Leu105 Leu102 Leu102 Leu103
Leu127 Leu143 Leu143 Leu146 Leu143 Leu143 Leu144
Ile131 Ile147 Ile147 Ile150 Leu147 Leu147 Ile148
Leu135 Leu151 Leu151 Leu154 Leu151 Leu151 Leu152
Ile139 Val155 Val155 Val158 Ile155 Ile155 Leu156
Leu153 Leu172 Leu172 Leu176 Leu171 Leu172 Leu170
Phe155 Phe174 Phe174 Phe178 Phe173 Phe174 Phe172
Phe157 Phe176 Phe176 Phe180 Phe175 Phe176 Phe174
Phe178 Phe197 Phe197 Phe201 Phe196 Phe197 Asn186
Leu192 Leu211 Leu211 Leu215 Leu210 Leu211 Met200
Ile196 Leu215 Met215 Met219 Leu214 Leu215 Leu204
Catalytic residues Lys176 Lys195 cons cons cons cons Leu
Asp211 Asp230 cons cons cons cons Asn
Supplemental Table 1 Continued
YHK2 HXK1 HXK2 HXK3 HKL1 HKL2 HKL3
Glucose contacts Ser158 Ser177 cons cons cons cons Thr
Asn237 Asn256 cons cons cons cons cons
Glu269 Glu284 cons cons cons cons cons Glu302 Glu315 cons cons cons cons cons
Conserved Gly Gly76 Gly91 Gly91 Gly94 Gly91 Gly91 Gly92
Gly80 Gly95 Gly95 Gly98 Gly95 Gly95 Gly96
Gly88 Gly103 Gly103 Gly106 Gly103 Gly103 Arg104
Gly154 Gly173 Gly173 Gly177 Ala172 Ala173 Gly171
Gly233 Gly252 Gly252 Gly256 Gly251 Gly252 Gly252
Gly235 Gly254 Gly254 Gly258 Gly253 Gly254 Gly254
Gly297 Gly310 Gly310 Gly312 Asn309 Asn310 Gly310
Gly307 Gly320 Gly320 Gly322 Gly319 Gly320 Gly320
Gly418 Gly440 Gly440 Gly441 Gly447 Gly444 Gly435
Gly461 Gly479 Gly479 Gly480 Ser486 Ser483 Ala474
Amino acid sequences were aligned using ClustalW in BioEdit. Annotations are based on regions homologous to yeast hexokinase II (Kuser et al. 2000). Designation of structural motifs as conserved (= cons), not conserved, or uncertain (uncer) is based on visual inspection and a relative assessment of the number of amino acid changes. However, the functional significance for a given motif change is not generally known such that the score of some motifs has some inherent ambiguity. For example, Loop 1 is LGGTN. This is changed in HKL1 to LGGTY and scored as uncertain with regard to whether this modified loop would still have similar functional capability. However, Loop 1 is more extensively changed in HKL3 to LRGKE and is scored as not conserved. Loop 2 in YHK2 is PDAMRTTQNP. The corresponding region in AtHXK1 is SIPPHLMTGG, but the function might still be retained even though the sequence is completely different. Loop 3 in YHK2 is SFPPASQ, which is present in most of the HXKs as SFPVKQ. This similarity was scored as conserved, except for the much more divergent sequence in HKL3 (TRSVEQ). The core sugar binding motif in YHK2 is LGFTFSF---Q--I, but in AtHXKs the conserved motif is LGFTFSF--Q--L/I. Both HKL1 and HKL2 were scored as having uncertain conservation of the core sugar sequence motif. HKL1 has a sequence of LAFTFSF------I, while HKL2 has a less divergent sequence of LAFTFSF---Q--I. YHK2 = yeast hexokinase 2; HXK1 = At4g29130; HXK2 = At2g19860; HXK3 = At1g47840; HKL1 = At1g50460; HKL2 = At3g20040; HKL3 = At4g37840.