TY - JOUR
T1 - A study on the conversion of procollagen
T2 - release and recovery of procollagen peptides in the culture medium
AU - Pontz, B. F.
AU - Muller, P. K.
AU - Meigel, W. N.
PY - 1973
Y1 - 1973
N2 - An in vitro study of the conversion of procollagen to collagen was carried out by culturing embryonic chicken calvaria and labeling them with [3,4 3H]proline and [35S]cysteine. A pulse experiment in the presence of α,α' dipyridyl, an iron chelator, was used to accumulate procollagen in the tissue. The presence of procollagen was demonstrated by the incorporation of [35S]cysteine and by its chromatographic behavior on CM cellulose under denaturing conditions. Using a parallel sample for a subsequent chase experiment in the absence of α,α' dipyridyl, only traces of procollagen were found in the tissue, since [35S]cysteine had disappeared and radioactive material chromatographed with the authentic α chains of lathyritic rat skin collagen. It is proposed that the conversion of procollagen to collagen is paralleled by the release of procollagen peptides into the culture medium, indicating a single step enzymatic cleavage. These peptides were isolated from the culture medium, and their molecular weights were determined as 18,000 for a [35S]cysteine containing and 12,000 for a [35S]cysteine free peptide. Both values are within the size range expected for peptides cleaved from procollagen chains of molecular weights between 110,000 and 120,000.
AB - An in vitro study of the conversion of procollagen to collagen was carried out by culturing embryonic chicken calvaria and labeling them with [3,4 3H]proline and [35S]cysteine. A pulse experiment in the presence of α,α' dipyridyl, an iron chelator, was used to accumulate procollagen in the tissue. The presence of procollagen was demonstrated by the incorporation of [35S]cysteine and by its chromatographic behavior on CM cellulose under denaturing conditions. Using a parallel sample for a subsequent chase experiment in the absence of α,α' dipyridyl, only traces of procollagen were found in the tissue, since [35S]cysteine had disappeared and radioactive material chromatographed with the authentic α chains of lathyritic rat skin collagen. It is proposed that the conversion of procollagen to collagen is paralleled by the release of procollagen peptides into the culture medium, indicating a single step enzymatic cleavage. These peptides were isolated from the culture medium, and their molecular weights were determined as 18,000 for a [35S]cysteine containing and 12,000 for a [35S]cysteine free peptide. Both values are within the size range expected for peptides cleaved from procollagen chains of molecular weights between 110,000 and 120,000.
UR - http://www.scopus.com/inward/record.url?scp=0015814594&partnerID=8YFLogxK
M3 - Article
C2 - 4745782
AN - SCOPUS:0015814594
SN - 0021-9258
VL - 248
SP - 7558
EP - 7564
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 21
ER -