In vitro synthesis of low molecular weight proteins in human platelets: absence of labelled release products.
Isolated human blood platelets incubated at 37 degrees C in vitro incorporated labelled amino acids into compounds which included some low molecular weight (less than 80KDa) proteins, as determined by autoradiography after sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions. Non-dialysable plasma factor(s) inhibited both uptake and incorporation, which although unaffected by Actinomycin D, was inhibited partly by Chloramphenicol and almost completely by Puromycin and Cycloheximide, results which confirm that synthesis is directed by pre-existing mRNAs, some of which is mitochondrial. Assuming that the mRNA coding for proteins which are truly "platelet specific" must be present in megakaryocyte cytoplasm, we investigated the possibility that such RNA may be sufficiently stable for its translation to continue in platelets. Although leakage from platelet alpha-granules and cytoplasm during incubation was negligible and platelets retained their secretory potential we were unable to detect radiolabelled proteins in thrombin-released material after incubation. We conclude that either alpha-granule proteins are not synthesised in platelets or their megakaryocyte progenitors, or that their mRNAs become degraded by the time platelets reach the peripheral circulation. Alternatively, the mechanism which concentrates these proteins in granules does not function in circulating platelets.
D000596:Amino Acids / Q000097:blood; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D001798:Blood Proteins / Q000096:biosynthesis*; D002701:Chloramphenicol / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D011691:Puromycin / Q000494:pharmacology; D012333:RNA, Messenger / Q000097:blood; D013917:Thrombin / Q000494:pharmacology
D000596:Amino Acids; D001798:Blood Proteins; D012333:RNA, Messenger; D011691:Puromycin; D002701:Chloramphenicol; D003513:Cycloheximide; D013917:Thrombin