Levels of sulfhydryls and disulfides in proteins from Neurospora crassa conidia and mycelia.
Proteins extracted with 6 M guanidine at 90 degrees C from conidia (asexual spores) of Neurospora crassa contained ca. 25% more total protein thiol and a fivefold-higher content of disulfide bonds than proteins extracted from mycelia, as determined by labeling with iodo[14C]acetic acid. The total thiol content was 88 mumol/g of protein in conidia and 70 mumol/g of protein in mycelia. The level of protein disulfide was 18.5 mumol/g of protein in conidia and 3.5 mumol/g of protein in mycelia, by the iodo[14C]acetic acid labeling method. Confirmatory results were obtained with 5'5-dithio-bis-2-nitrobenzoic acid titration of protein thiol groups in 1% sodium dodecyl sulfate as well as by amino acid analysis of cysteic acid derivatives. Buffer-extracted proteins from conidia, but not mycelia, were found to contain enriched levels of protein thiols and disulfides per gram of protein as compared with guanidine hydrochloride extracts. It was demonstrated that the high disulfide content of crude conidial extracts was not due to measurable levels of mixed disulfides formed between protein sulfhydryl groups and cysteine. During germination of the conidia, the high disulfide levels of the conidial proteins remained constant. These data suggest that, unlike the disulfides of glutathione, the bulk of conidial protein disulfides were not reduced, excreted, or extensively degraded during germination.
D000596:Amino Acids / Q000032:analysis; D002250:Carbon Radioisotopes; D004220:Disulfides / Q000032:analysis*; D005033:Ethylmaleimide; D005656:Fungal Proteins / Q000302:isolation & purification*; D007461:Iodoacetates; D019807:Iodoacetic Acid; D009491:Neurospora / Q000032:analysis*; D009492:Neurospora crassa / Q000032:analysis* / Q000254:growth & development; D013172:Spores, Fungal / Q000032:analysis; D013438:Sulfhydryl Compounds / Q000032:analysis*
D000596:Amino Acids; D002250:Carbon Radioisotopes; D004220:Disulfides; D005656:Fungal Proteins; D007461:Iodoacetates; D013438:Sulfhydryl Compounds; D005033:Ethylmaleimide; D019807:Iodoacetic Acid