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Table 1 Mass spectrometric identification of fibronectin-, vitronectin- and laminin-binding C. parapsilosis and C. tropicalis cell wall-associated proteins

From: Fibronectin-, vitronectin- and laminin-binding proteins at the cell walls of Candida parapsilosis and Candida tropicalis pathogenic yeasts

C. parapsilosis
NCBI protein database accession number Protein Molecular mass [kDa] Scores Number of peptides Sequence coverage [%] Method of extraction from fungal cell wall Method of identification
β-1,3-glucanase β-1,6-glucanase AFC CL
Fibronectin-binding proteins
gi|354547941 hypothetical protein CPAR2_404800 [C. parapsilosis], cell wall agglutinin, N-terminal, ligand sugar binding 143.6 736 13 14   + +  
gi|354547939 hypothetical protein CPAR2_404780 [C. parapsilosis], cell wall agglutinin, N-terminal, ligand sugar binding 113.6 525 10 13   + +  
gi|354546787 hypothetical protein CPAR2_211630 [C. parapsilosis], similar to C. albicans Eft2, elongation factor 2 93.8 259 11 14 + + + +
gi|354547813 hypothetical protein CPAR2_403510 [C. parapsilosis], similar to C. albicans Rbt1, cell wall protein with similarity to Hwp1, required for virulence 83.8 73 2 1   +   +
gi|354545198 hypothetical protein CPAR2_804740 [C. parapsilosis], similar to C. albicans Tkl1, putative transketolase 74.6 166 6 8 + + + +
gi|354545113 hypothetical protein CPAR2_803890 [C. parapsilosis], similar to C. albicans Mls1, malate synthase 62.7 1434 32 72 + + + +
gi|354544475 hypothetical protein CPAR2_301880 [C. parapsilosis], putative amidase 61.7 938 18 38 + + + +
gi|354546116 hypothetical protein CPAR2_204880 [C. parapsilosis], similar to C. albicans Pgi1, putative glucose-6-phosphate isomerase 61.5 104 3 7   + +  
gi|354545980 hypothetical protein CPAR2_203520 [C. parapsilosis], similar to C. albicans Gnd1, putative 6-phosphogluconate dehydrogenase 55.4 184 5 15 + +   +
gi|354546805 hypothetical protein CPAR2_211810 [C. parapsilosis], similar to C. albicans Gpm1, phosphoglycerate mutase 27.6 366 13 33   +   +
Vitronectin-binding proteins
gi|8927048 elongation factor 2 [C. parapsilosis] 90.3 362 8 10 + + + +
gi|354545198 hypothetical protein CPAR2_804740 [C. parapsilosis], similar to C. albicans Tkl1, putative transketolase 74.6 146 3 4 + + + +
gi|354545113 hypothetical protein CPAR2_803890 [C. parapsilosis], similar to C. albicans Mls1, malate synthase 62.7 927 19 43   + + +
gi|354544475 hypothetical protein CPAR2_301880 [C. parapsilosis], putative amidase 62.1 1452 25 45 + + + +
gi|354546116 hypothetical protein CPAR2_204880 [C. parapsilosis], similar to C. albicans Pgi1, putative glucose-6-phosphate isomerase 61.5 655 15 22   + +  
gi|354545980 hypothetical protein CPAR2_203520 [C. parapsilosis], similar to C. albicans Gnd1, putative 6-phosphogluconate dehydrogenase 55.4 636 14 31 + + + +
gi|354546348 hypothetical protein CPAR2_207210 [C. parapsilosis], similar to C. albicans Eno1, enolase 47 229 4 13   + +  
gi|354545888 hypothetical protein CPAR2_202600 [C. parapsilosis], similar to C. albicans Tal1, putative transaldolase 35.7 526 10 36   + +  
Laminin-binding proteins
gi|354547939 hypothetical protein CPAR2_404780 [C. parapsilosis], cell wall agglutinin, N-terminal, ligand sugar binding 113.6 93 3 2   + +  
gi|354545113 hypothetical protein CPAR2_803890 [C. parapsilosis], similar to C. albicans Mls1, malate synthase 62.7 425 9 20   +   +
gi|354544475 hypothetical protein CPAR2_301880 [C. parapsilosis], putative amidase 61.7 1629 67 58 + +   +
gi|354546805 hypothetical protein CPAR2_211810 [C. parapsilosis], similar to C. albicans Gpm1, phosphoglycerate mutase 27.3 167 4 18 +    +
Fibronectin-binding proteins
gi|255722852 predicted protein [C. tropicalis MYA-3404], hyphally regulated cell wall protein, N-terminal 194.6 202 6 4   + +  
gi|255728333 malate synthase [C. tropicalis MYA-3404] 62.4 921 23 58   + + +
gi|255727881 enolase 1 [C. tropicalis MYA-3404] 46.9 619 33 55   + + +
gi|255732698 fructose-1,6-bisphosphatase [C. tropicalis MYA-3404] 40.5 134 4 14 + + + +
gi|255722021 transaldolase [C. tropicalis MYA-3404] 35.5 123 3 8   + +  
Vitronectin-binding proteins
gi|255722852 predicted protein [C. tropicalis MYA-3404], hyphally regulated cell wall protein, N-terminal 194.6 107 3 2   + +  
gi|240134900 elongation factor 2 [Candida tropicalis MYA-3404] 90.2 378 10 11   + +  
gi|255728333 malate synthase [C. tropicalis MYA-3404] 62.4 529 12 25   +   +
gi|255727881 enolase 1 [C. tropicalis MYA-3404] 46.9 157 5 18 +   +  
gi|255732698 fructose-1,6-bisphosphatase [C. tropicalis MYA-3404] 40.5 320 7 25 + + + +
gi|255722021 transaldolase [C. tropicalis MYA-3404] 35.5 513 10 28   +   +
Laminin-binding proteins
gi|255722852 predicted protein [C. tropicalis MYA-3404], hyphally regulated cell wall protein, N-terminal 194.6 103 2 1   + +  
gi|240134900 elongation factor 2 [Candida tropicalis MYA-3404] 90.2 259 7 8   + +  
gi|255728333 malate synthase [C. tropicalis MYA-3404] 62.4 337 10 22   + + +
gi|240132975 peroxisomal catalase [Candida tropicalis MYA-3404] 55.1 104 3 5   + +  
gi|255727881 enolase 1 [C. tropicalis MYA-3404] 46.9 189 4 12   + +  
gi|255732698 fructose-1,6-bisphosphatase [C. tropicalis MYA-3404] 40.5 136 4 15 +   +  
gi|255722021 transaldolase [C. tropicalis MYA-3404] 35.5 487 12 44   + +  
  1. Cell wall-associated proteins were extracted from C. parapsilosis and C. tropicalis pseudohyphae, using either β-1,3-glucanase (+β-mercaptoethanol) or β-1,6-glucanase. Subsets of ECMP-binding proteins were isolated from the whole cell wall protein mixtures, using affinity chromatography (AFC) or cross-linking (CL) methods. After SDS-PAGE electrophoresis, specific protein bands were excised and digested using trypsin. Peptides were analyzed using the UltiMate 3000 RSLCnano System coupled to a micrOTOF-QII mass spectrometer and Apollo Source ESI nano-sprayer (for AFC-isolated proteins) or an Dionex Ultimate 3000 UHPLC system coupled to an HCT Ultra ETDII mass spectrometer equipped with an ESI ion source (for CL-isolated proteins). The obtained lists of peaks were searched against the NCBI protein database using an in-house Mascot server. The data represent a combination of results obtained for the extracts of cell wall-associated proteins, prepared from three independent yeast cultures. For a given extraction method (β-1,3-glucanase vs. β-1,6-glucanase) and ECMP-binder isolation method (AFC vs. CL), only those proteins are listed that were found on the analysis of all three yeast cultures. Hence, each “+” sign means that a given protein was found at least three times. The best Mascot parameters (scores, number of identified peptides and sequence coverage) ever recorded for a given protein are specified; the proteins identified with a score > 70 and a number of identified peptides ≥ 2 are only listed