Table 2.
Proteins identified by LC‐MS/MS from spots that differed in their expression ratio among growth of wild type (w.t.) of Colletotrichum acutatum under nitrogen‐limiting conditions (mm), w.t. growth under complete nutrient supply (reg) and during the appressorium formation stage (app). Proteins were classified according to functional categories
| Function * | Spot† | GenBank accession no.‡ | Best database match§ | Mol. mass;pI/ ¶ experimental/ theoretical | Coverage (%)†† | Total score‡‡ | App Reg MM regulation§§ |
|---|---|---|---|---|---|---|---|
| ROS scavenging and stress response | 4 | AAD19338 | Cu‐Zn superoxide dismutase (Glomerella cingulata) | 20.3;6.3/16;5.9 | 29(8) | 90 | +–+ |
| 14 | EAA74714 | Hypothetical protein, similar to gluthatione peroxidase (Gibberella zeae) | 18.8;6.5/19.1;6.3 | 15(5) | 93.3 | +–+ | |
| 15 | EAA57651 | Hypothetical protein, CD: pfam00012 Hsp70 protein (Aspergillus nidulans) | 26.3;5.8/72.3;5.7 | 8(4) | 90 | +–– | |
| 33 | EAA77628 | Hypothetical protein, similar to putative stress response protein (Gibberella zeae) | 20.6;4.7/28.4;4.33 | 9(2) | 82 | –++ | |
| 40 | XP366662 | Hypothetical protein, similar to cell death‐related protein (Magnaporthe grisea) | 15.9;5.1/15;5.3 | 8(1) | 93 | ––+ | |
| 60 | AP007175.1 | Hypothetical protein, similar to bifunctional catalase peroxidase (Cat2) (Aspergillus oryzae) | 80;6.2/82;5.8 | 4(4) | 91 | +–– | |
| Main carbohydrate, glyoxylate and lipid metabolism | 1 | EAA59740 | Hypothetical protein, CD: Cytochrome c oxidase subunit pfam02284 (Aspergillus nidulans) | 14.8;4.8/17.8.5;5.6 | 10(2) | 93 | –++ |
| 8 | CAF31997 | Putative isocitrate dehydrogenase (Aspergillus fumigatus) | 40.4;4.1/41.7;8.7 | 10(2) | 97 | ndi+– | |
| 20 | CAA82232 | Enolase 2 (Ricinus communis) | 25.9;6.14/47.9;5.6 | 13(3) | 90.6 | –++ | |
| 36 | XP680863 | Hypothetical protein, glutathione‐ dependent formaldehyde activating (Aspergillus nidulans) | 14;6.7/14.7;6.03 | 17(5) | 92 | +–– | |
| 37 | P54117 | Glyceraldehyde 3‐phosphate dehydrogenase (Glomerella lindemuthiana) | 24.18;6.6/36.3;6.54 | 12(4) | 85 | –++ | |
| 44 | EAA69530 | Hypothetical protein, CD: pfam00171 aldehyde dehydrogenase family (Gibberella zeae) | 53.6;6.18/53.4;5.4 | 9(4) | 92 | +–– | |
| 52 | EAA75069 | Hypothetical protein, CD: Formate dehydrogenase NAD‐ dependent (Gibberella zeae) | 38.6;6.2/40;6.1 | 16(8) | 94 | +–– | |
| 53 | EAA68236 | Hypothetical protein, CD: Malate dehydrogenase (Gibberella zeae) | 23.7;6.5/34;6.4 | 25(8) | 93 | +–– | |
| 56 | EAA73569 | Hypothetical protein, CD: Acetyl‐CoA acetyltransferase (Gibberella zeae) | 49.6;6.6/43.6;7.5 | 6(3) | 83 | +–+ | |
| 58 | EAA77220 | Hypothetical protein, similar to succinate dehydrogenase (Gibberella zeae) | 196.7;6/50.2;5.8 | 1(1) | 92 | ––– | |
| Cellular, signal transduction and co‐factor metabolism | 2 | Q7RVL0 | GTP‐binding nuclear protein GSP1/Ran (Neurospora crassa) | 14.8;5/24.2;6.4 | 21(5) | 93 | nd nd + |
| 7 | XP457640 | Hypothetical protein, similar to CipC protein of Emericella nidulans (Debaryomyces hansenii) | 14;5.9/14.9;5.3 | 14(4) | 86 | +–– | |
| 26 | XP328263 | Hypothetical protein, similar to protoporphyrinogen oxidase [Coenzyme metabolism] (Neurospora crassa) | 60;6.17/57.9;5.8 | 8(3) | 83 | +–– | |
| 28 | S30118 | Orotate phosphoribosyl transferase (Colletotrichum graminicola) | 28.7;6.7/25.2;5.9 | 11(2) | 89 | +–– | |
| 42 | XP326472 | Hypothetical protein, CD: Calmodulin‐related Ca2+‐binding protein (Neurospora crassa) | 13.6;4.3/16.3;4.5 | 18(2) | –++ | ||
| 48 | EAA74730 | Hypothetical protein, similar to secretory pathway gdp dissociation inhibitor CD: pfam00996 (Gibberella zeae) | 54.5;5.6/51.4;5.37 | 9(4) | 94 | +–– | |
| 49 | CAD60606 | Hypothetical protein, CD: Thiosulfate sulfur transferase (Podospora anserine) | 35.6;6/38.4;6.7 | 10(3) | 91 | +–+ | |
| 55 | EAA76084 | Hypothetical protein, CD: pfam00890 FAD binding domain, Cytochrome b5‐like heme/steroid binding domain (Gibberella zeae) | 70.2;6.6/67.9;6 | 16(13) | 88 | +–– | |
| 57 | XP323373 | b‐tubulin chain (Neurospora crassa) | 38.5;5.2/49.4;4.84 | 8(3) | 85 | –++ | |
| Nitrogen and amino acid metabolism. Protein synthesis, transcription and degradation | 5 | XP322910 | Ketol‐acid reductoisomerase precursor (Neurospora crassa) | 29.2;6.5/44.6;8.5 | 6(2) | 95 | ––+ |
| 16 | AAB00322 | Glutamine synthetase (Glomerella cingulata) | 39.9;5.7/39.9;5.7 | 23(11) | 89 | +–+ | |
| 22 | CAD70393 | Conserved hypothetical protein, similar to 20S proteasome subunit alpha type 6 (Neurospora crassa) | 29.5;6/27.7;5.6 | 13(4) | 90 | ––+ | |
| 32 | BAE62886 | Hypothetical protein, similar to 26s proteasome regulatory subunit mts4 (Aspergillus oryzae) | 41.4;6.7/100;4.89 | 1(1) | 89 | ++– | |
| 34 | AAF82115 | Cobalamin‐independent methionine synthase (Aspergillus nidulans) | 14.9;5.9/86.8;6.36 | 3(3) | 93 | ––+ | |
| 39 | EAA77131 | EF2_NEUCR elongation factor 2 (EF‐2) (Gibberella zeae) | 41.5;6.2/91.6;6.45 | 6(4) | 95 | +–+ | |
| 41 | CAC28704 | Hypothetical protein, similar to ubiquitin‐conjugating enzyme ubcP3 (Neurospora crassa) | 16.8;5.2/18.5;5 | 5(1) | 93 | –++ | |
| 43 | EAA68262 | Hypothetical protein, similar to prolidase (Gibberella zeae) | 21.4;6.2/26.1;5.8 | 6(2) | 86 | +–– | |
| 46 | CAF05873 | Glycine hydroxymethyl transferase cytosolic (Neurospora crassa) | 23.7;6.5/52.9;6.93 | 6(2) | 82 | +–– | |
| 51 | XP_682383 | Hypothetical protein, similar to threonine dehydrogenase and related Zn‐dependent dehydrogenases (Aspergillus nidulans) | 40.7;6.2/42.5;5.3 | 8(3) | 89 | +–– | |
| Unknown function | 38 | XP328441 | Hypothetical protein (Neurospora crassa) | 26.5;6/36.2;6.3 | 8(2) | 90 | nd –+ |
Main functional categories.
Assigned spot number as indicated in Fig. 1.
Accession numbers in the NCBI databases.
Best database match according to NR‐NCBI or specific databases; in the case of hypothetical proteins, putative functions were found by using the on‐line conserved domains (CD) search in the NCBI database. The CD with the highest score was listed as the CD for the respective hypothetical protein. The corresponding organism from which the identified protein originates is denoted in parentheses.
Experimental and theoretical mass (kDa) and pI of identified protein. Experimental values were calculated with PD‐Quest software and standard molecular‐weight markers. Theoretical values were retrieved from the protein database.
Percentage sequence coverage and number of peptides identified in parentheses.
Total peptide score: a peptide was considered to be of high quality if its Pep‐Miner identification score was greater than 80 and had a sequest Xcore of 1.5 for single‐charged peptides, 2.5 for double‐charged peptides and 3 for triple‐charged peptides.
Significantly up‐ or down‐regulated (+/–, respectively, among treatments).
nd, not detectable.