Extracellular labeling of growing secreted polypeptide chains in Bacillus subtilis with diazoiodosulfanilic acid.
Studies of the mechanism of protein secretion in a Gram-positive bacterium, Bacillus subtilis, yielded results very similar to those previously obtained with a Gram-negative organism: nascent chains protruding from protoplasts could be labeled extracellularly; the labeled chains could be recovered on polysomes isolated from the membrane--polysome fraction; they could be released by puromycin, low Mg2+, or chain completion; the completed chains include a known secreted protein (alpha-amylase); and their ribosomes appear to be attached to membrane solely by their nascent chains. The reagent used for extracellular labeling, [1252]diazoiodosulfanilic acid, yielded severalfold more specific labeling of the nascent chains (7--10% of the total cellular labeling and one-fourth to one-third of that of the membrane--polysome fraction) than was obtained earlier with another nonpenetrating reagent.
D000681:Amylases / Q000096:biosynthesis*; D001391:Azo Compounds; D001412:Bacillus subtilis / Q000378:metabolism*; D003979:Diazonium Compounds; D009829:Oleic Acids / Q000378:metabolism; D010452:Peptide Biosynthesis; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D011523:Protoplasts / Q000378:metabolism; D011691:Puromycin / Q000494:pharmacology; D012260:Ribonucleases; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism; D013425:Sulfanilic Acids / Q000031:analogs & derivatives; D014498:Uracil / Q000378:metabolism; D000516:alpha-Amylases / Q000096:biosynthesis*
D001391:Azo Compounds; D003979:Diazonium Compounds; D009829:Oleic Acids; D013425:Sulfanilic Acids; D011691:Puromycin; D014498:Uracil; C018909:diazoiodosulfanilic acid; D012260:Ribonucleases; D000681:Amylases; D000516:alpha-Amylases