Supplementary Table 1-Results of mass spectrometric analysis of Cryptosporidium protein spots selected from 2-DGE
Cy5 dye labeled Cryptosporidium sporozoites membrane proteins after interaction with the host cells (HCT-8 cells)
S. No. / PlasmodiumAccession No. / Mascot Score / Cryptosporidium
Accession No. / Cryptosporidium
Protein
name / Query coverage / Max. identity / References for Cryptosporidium / Cellular locations / Putative functions
gi|2281619
merozoite surface protein 3 / 21 / AAF81669.1 / glycoprotein 15 kDa / 21% / 35% / Jenkins et al. 1993;
Cevallos et al. 2000;
Strong et al. 2000 / surface of sporozoites andmerozoites / adherence and invasion
gi|2281619 / 21 / AAF81669.1 / glycoprotein 40/15 kDa / 21% / 35% / Jenkins et al. 1993;
Cevallos et al. 2000 / apical end of sporozoites,
sporozoites surface / adherence and invasion
gi|81176680
TRAP type 1 / 20 / ACX31742.1 / TRAP- C1 / 19% / 73% / Spano et al. 1998 / apical complex of sporozoites, probably in micronemes / adhesion and locomotion
20 / CAA54689.1 / TRAP-C2 / 59% / 40% / Spano et al. 1998 / apical complex of sporozoites,
probably in micronemes / adhesion and locomotion
20 / XP_626502.1 / CpTSP10 / 64% / 28% / Putignani et al. 2008 / apical complex of sporozoites and merozoites / adhesive protein
secreted by sporozoites during gliding motility
gi|78032663
erythrocyte membrane protein (EMP1) / 26 / AAC98153.1 / glycoprotein 900 kDa / 54% / 26% / Petersen et al. 1992;
Barnes et al. 1998 / micronemes of sporozoites and merozoites.
Sporozoites surface / adherence and invasion
gi|224813714
EMP1 / 21 / XP_626655.1 / mucin / 21% / 38% / O’Connor et al. 2009 / micronemes of sporozoites and merozoites.
Sporozoites surface / adherence and invasion
gi|323393297
EMP1 / 24 / XP_625734.1 / ABC transporter protein / 77% / 28% / Perkins et al. 1999 / host parasite boundry, Parasitophorous
vacuole / nutrients transport
gi|224813676
EMP1 / 24 / XP_627856.1 / zinc protease / 51% / 31% / Okhuysen et al.1996 / NA / excystation
24 / XP_628686.1 / V -ATPase / 24% / 29% / Abrahamsen et al. 2004 / NA / lipids transport
gi|78040983
EMP1 / 8 / XP_627588.1 / CpTSP6 / 61% / 32% / Putignani et al. 2008 / apical complex of sporozoites and merozoites / adhesive protein
secreted by sporozoites during gliding motility
gi|40646474
EMP1 / 26 / XP_627454.1 / CpTSP9 / 31% / 27% / Putignani et al. 2008 / apical complex of sporozoites and merozoites / adhesive protein
secreted by sporozoites during gliding motility
gi|323393297
EMP1 / 6 / XP_626895.1 / Hsp70 / 28% / 19% / Khramtsov et al. 1995 / NA / stress protection
gi|197252799
EMP1 / 23 / XP_625628.1 / Hsp90 / 20% / 43% / Woods et al. 1999 / NA / stress protection
gi|31322987
var2csa / 6 / AAN60755.1 / polyketide synthase / 44% / 27% / Zhu et al. 2002 / NA / virulence factor with unknown function
gi|113242
Actin-2 / 84 / XP_001388282.1 / actin / 99% / 82% / Wetzel et al. 2005 / cytoskeleton / glinding motility
gi|124505225 / 7 / XP_627811.1 / serine protease / 19% / 40% / Forney et al. 1996 / NA / excystation
7 / CAD98301.1 / subtilisin-like serine protease / 19% / 40% / Forney et al. 1996 / NA / excystation
gi|124506241
cation-transporting ATPase 1 / 33 / XP_001388220.1 / ATPase 2 / 18% / 24% / LaGier et al. 2001 / parasitophorous vacuole membrane / biomembrane heavy metal Transporter,
gi|237780847
var DBLalpha / 25 / XP_626689.1 / P-ATPase / 11% / 62% / Zhu and Keithly. 1997 / parasitophorous vacuole membrane / lipid transport
gi|258597280
conserved Plasmodium protein / 7 / XP_628694.1 / secreted protein / 43% / 33% / Abrahamsen et al. 2004 / NA / host cell invasion
gi|1084996
Plasmepsin II / 23 / XP_625608.1 / CpTSP7 / 65% / 71% / Putignani et al. 2008 / apical complex of sporozoites and merozoites / adhesive protein
secreted by sporozoites during gliding motility
24 / XP_626191.1 / WD repeat protein / 45% / 75% / Trasarti et al. 2007 / NA / protein-protein interactions
21 / XP_627787.1 / myosin / 12% / 47% / Wetzel et al. 2005 / cytoskeleton / gliding motility
23 / CAD98380.1 / PI3 kinase / 60% / 100% / Nelson et al. 2006 / NA / involved in cell signaling
gi|124802756
calmodulin, / 10 / XP_001388192.1 / calmodulin / 67% / 27% / NR / cytoskeleton / gliding motility
gi|81176680
TRAP type 1 / 20 / CAD98628.1 / sporozoite protein / 64% / 37% / Riggs et al. 1997 / Sporozoites surface / adherence and invasion
gi|124801145
conserved Plasmodium protein / 21 / XP_626874.1 / Ser/thr protein kinase / 72% / 31% / NR / NA / phosphorylation and probably in cell signaling
gi|124506803
conserved Plasmodium protein, / 8 / XP_001388075.1 / Hsp 105kD / 27% / 43% / NR / NA / may involve in stress protection with unknown function
gi|258597009
ankyrin-repeat protein / 20 / CAD98264.1 / ankyrin-repeat protein / 75% / 34% / NR / NA / may involve in protein- protein interaction with unknown function
gi|1084996
Plasmepsin II / 23 / CAD98380.1 / PI4kinase / 60% / 100% / NR / NA / probably in cell signaling
23 / XP_627059.1 / coiled coil protein / 30% / 83% / NR / NA / Functionally uncharacterized. No conserved domain found
23 / XP_628582.1 / signal peptide, large secreted protein / 70% / 80% / NR / NA / Functionally uncharacterized. May involve in host cell invasion.
23 / XP_626282.1 / transport protein / 20% / 100% / NR / NA / Functionally unchara-cterized but conserved domain search shows
ABC transporter. Thus, probably involved in nutrients transport
23 / XP_626864.1 / 12 transmembrane domain protein / 45% / 67% / NR / Membrane / Functionally uncharacterized but conserved domain search shows
MFS family sugar transporter. Therefore, may involve in nutrients transport.
23 / XP_001388185.1 / hypothetical protein / 35% / 100% / NR / NA / Functionally unchara-cterized but conserved domain search shows
ABC transporter So, possibly may involve in nutrients transport
23 / XP_625300.1 / GMP synthase / 25% / 80% / NR / membrane / Functionally unchara-cterized but conserved domain search shows
amidotransferase activity
gi|323392810
EMP1 / 6 / XP_628430.1 / integral membrane protein with transmembrane domains / 27% / 26% / NR / membrane / Functionally uncharacterized. No conserved domain found
gi|197252993
EMP1 / 14 / XP_628061.1 / integral membrane protein / 26% / 38% / NR / membrane / Functionally uncharacterized. No conserved domain found
gi|124806346
conserved Plasmodium protein / 27 / CAD98374.1 / G-protein associated signal transduction protein / 48% / 32% / NR / membrane / Unknown function
gi|298716900
surfin 4.2 / 20 / XP_627036.1 / integral membrane protein / 11% / 36% / NR / membrane / Functionally uncharacterized. No conserved domain found
gi|1084996
Plasmepsin II / 23 / XP_626026.1 / protein with 5 transmembrane domains / 20% / 100% / NR / membrane / Functionally uncharacterized. No conserved domain found
24 / XP_628636.1 / membrane protein / 20% / 100% / NR / membrane / Functionally uncharacterized. No conserved domain found
23 / XP_627684.1 / membrane associated protein with 2 transmembrane domains / 60% / 100% / NR / membrane / Functionally uncharacterized. No conserved domain found
gi|224813676
EMP1 / 24 / XP_627924.1 / 9 trans
membrane domain protein / 30% / 30% / NR / membrane / Functionally uncharacterized. No conserved domain found
gi|1084996
Plasmepsin II / 23 / XP_627268.1 / membrane associated protein, signal peptide, transmembrane domain / 35% / 100% / NR / membrane / Functionally uncharacterized. No conserved domain found
23 / CAD98664.1 / hypothetical protein / 30% / 83% / NR / NA / Functionally uncharacterized. No conserved domain found.
23 / XP_627628.1 / hypothetical protein / 35% / 100% / NR / NA / Functionally uncharacterized. No conserved domain found.
23 / XP_625663.1 / hypothetical protein / 80% / 100% / NR / NA / Functionally uncharacterized. No conserved domain found.
23 / XP_627456.1 / hypothetical protein / 45% / 100% / NR / NA / Functionally uncharacterized. No conserved domain found.
23 / XP_625471.1 / hypothetical protein / 20% / 100% / NR / NA / Functionally uncharacterized. No conserved domain found.
23 / XP_001388143.1 / hypothetical protein / 35% / 100% / NR / NA / Functionally uncharacterized.
23 / XP_001388051.1 / hypothetical protein / 90% / 100% / NR / NA / Functionally uncharacterized.
23 / XP_627097.1 / hypothetical protein / 85% / 100% / NR / NA / Functionally uncharacterized.
gi|17148495 / 23 / XP_625438.1 / hypothetical protein / 81% / 41% / NR / NA / NA
gi|1084996 / 23 / XP_627628.1 / hypothetical protein / 35% / 100% / NR / NA / NA
gi|224813676
EMP1 / 24 / XP_626096.1 / pre-mRNA splicing factor / 25% / 34% / - / - / -
gi|552237
ubiquitin / 11 / XP_626185.1 / ubiquitin domain at the C-terminus / 41% / 25% / - / - / -
gi|197252993
EMP1 / 28 / XP_627569.1 / CpCOWP1, oocyst wall protein / 26% / 32% / - / - / -
gi|82658995
EMP1 / 23 / XP_627568.1 / Kem1p-like 5'-3' exonuclease / 20% / 33% / - / - / -
gi|40646474
EMP1 / 26 / XP_626800.1 / nucleoporin, FG-rich motifs / 19% / 46% / - / - / -
*NA-Not Applicable, NR-Not Reported
≠MS spectra were searched against NCBI nr database forPlasmodium protein database due to unavailability of Cryptosporidium database in the software. The results obtained were further subjected to protein BLAST against Cryptosporidiumdatabase to find homology with default parameters.
±Information regarding function of each identified proteins were extracted from published literature as well as from protein database (
¥ TRAP - Thrombospondin (TSP)-related adhesive protein
ABC transporter protein - ATP-binding cassette transporter protein
ATPase 2 - Adenosine triphosphatase 2
P-ATPase - Phospholipid translocating ATPase
V –ATPase - Vacuolar proton translocating ATPase
CpTSP6 - Thrombospondin-related proteins
Hsp - Heat shock proteins
PI3_PI4_kinase - Phosphatidylinositol-3 phosphatidylinositol-4 kinase
GMP synthase - Guanine monophosphate synthetase
γ pre-mRNA splicing factor, nucleoporin FG-rich motifs, Kem1p-like 5'-3' exonuclease, ubiquitin domain at the C-terminus, CpCOWP1 Cryptosporidium oocyst wall protein- nonspecific proteins.
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