Evidence for the synthesis of pro-CRF (corticotropin-releasing factor) in cultured fetal pancreatic islets of rats.
Immunoreactive species extracted from cultured pancreatic fetal islets of rat have been immunoprecipitated with anti-CRF (corticotropin-releasing factor) antibody and labeled with 125I. This material was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These immunoreactive species corresponded to CRF-like material detected by immunocytochemistry in certain cells of the islets of Langerhans. The analysis of immunoprecipitated material showed that it contained an immunoreactive polypeptide chain of 17,000. This band corresponds to the precursor minus the signal peptide as deduced by cDNA cloning analysis. A lower molecular weight immunoreactive material was also detected, apparently derived from the precursor by peptide bond cleavage which yielded intermediate fragments. These bands disappeared and generated a Mr 4000 when the corresponding species, fractionated by Sephadex G-25 column, were incubated with isolated islet extracts at pH 5.5. In isolated islets from 3-day-old rat, only the Mr 17,000 and 4000 species appeared. These results suggest strongly that CRF is synthesized in situ and that the conversion process could involve a sequential rather than a single cleavage of pro-CRF. The major end product was probably CRF, whereas intermediate forms seem to carry an NH2 terminal extension of CRF.
Robichon
A
A
Laboratoire de Bioactivation des Peptides, Institut Jacques Monod, Faculté de Jussieu, Paris, France.
Nicolas
P
P
D000818:Animals; D002478:Cells, Cultured; D003346:Corticotropin-Releasing Hormone / Q000096:biosynthesis*; D005260:Female; D005333:Fetus / Q000166:cytology / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D011247:Pregnancy; D011498:Protein Precursors / Q000096:biosynthesis*; D051381:Rats; D011919:Rats, Inbred Strains
D011498:Protein Precursors; C041645:pro-corticotropin releasing hormone; D003346:Corticotropin-Releasing Hormone