Isolation and characterization of collagen-synthesizing polysomes from chick embryos.
Collagen-synthesizing polysomes were isolated by low-speed centrifugation of the post-mitochondrial supernatant of chick homogenates. Electron microscopy of the fraction thus isolated shows it to be exclusively composed of ribosomes. Amino acid incorporation in vitro showed that these particles were efficient in the incorporation of proline, but not tryptophan, as opposed to ribosomes obtained from the supernatant of the low-speed centrifugation. The incorporation process was highly dependent on GTP, and exibited an optimal Mg2+concentration of 5.6mM. The reaction was inhibited by RNase, elongation inhibitors as anysomycin, sparsomycin, fusidic acid and GDPCP. It was also moderately inhibited by initiation inhibitors such as aurintricarboxilic acid and pyrocatechol violet. The product of the incorporation was characterized as collagen by its sensitivity towards purified collagenase, lack of tryptophan, chromatography in CM-cellulose and molecular sieve chromatography in Sephadex G-200.
D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D002458:Cell Fractionation; D002642:Chick Embryo; D003094:Collagen / Q000096:biosynthesis*; D005672:Fusidic Acid / Q000494:pharmacology; D006150:Guanine Nucleotides / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D003012:Microbial Collagenase; D008928:Mitochondria / Q000378:metabolism; D008970:Molecular Weight; D011132:Polyribosomes / Q000378:metabolism* / Q000648:ultrastructure; D011392:Proline / Q000378:metabolism; D012260:Ribonucleases / Q000494:pharmacology; D012270:Ribosomes / Q000378:metabolism; D013033:Sparsomycin / Q000494:pharmacology; D014364:Tryptophan / Q000378:metabolism
D006150:Guanine Nucleotides; D005672:Fusidic Acid; D000841:Anisomycin; D013033:Sparsomycin; D014364:Tryptophan; D003094:Collagen; D011392:Proline; D012260:Ribonucleases; D003012:Microbial Collagenase; D008274:Magnesium