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. 1979 Jul 15;182(1):81–93. doi: 10.1042/bj1820081

Translation of embryonic-chick tendon procollagen messenger ribonucleic acid in two cell-free protein-synthesizing systems

Kathryn S E Cheah 1, Michael E Grant 1, David S Jackson 1
PMCID: PMC1161236  PMID: 227370

Abstract

Embryonic-chick tendon poly(A)-containing RNA was translated in the wheat-germ and mRNA-dependent rabbit reticulocyte-lysate systems. The ability of each system to synthesize polypeptides similar to pro-α chains of collagen was tested on the bases of electrophoretic mobility and susceptibility to highly purified bacterial collagenase. Very small amounts of polypeptides in the size range of pro-α chains were synthesized in the wheat-germ system, whereas efficient synthesis of two polypeptides similar to pro-α1 and pro-α2 chains was achieved in the reticulocyte lysate. The collagenous nature of the major high-molecular-weight products synthesized was demonstrated by their susceptibility to collagenase and ability to act as a substrate for purified collagen proline hydroxylase. Determinations of the relative amounts of these translation products suggest that the 2:1 ratio of pro-α1 and pro-α2 chains found in type I procollagen is reflected in proportional amounts of translatable mRNA for pro-α1 and pro-α2 chains. Comparisons of the electrophoretic mobilities of hydroxylated and unhydroxylated reticulocyte-lysate translation products were made with appropriate standards of hydroxylated and unhydroxylated procollagen polypeptides. The results suggest that, in common with a number of secreted proteins, procollagen is synthesized as pre-pro molecules consistent with the `Signal Hypothesis'.

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