Generation and characterization of an antiserum reactive with a proteolytic processing site within rat procorticotrophin-releasing hormone

Neuropeptides
M G CastroP R Lowenstein

Abstract

In this paper we report the generation of an antibody specific for the cleavage site within procorticotrophin-releasing hormone (proCRH) at the N-terminus proCRH/CRH (1-41) junction. Using radioimmunoassay techniques were show that the antibody generated (781) cross-reacts specifically with the proCRH (137-150) Tyr fragment, corresponding to the cleavage site within the full length precursor molecule. The anti-cleavage site antibody does not crossreact with the endoproteolytic products originated from the CRH precursor molecule, i.e. CRH (1-41) or proCRH (125-151) or with any of the CRH-immunoreactive fragments tested i.e. CRH (36-41), CRH (1-20) and CRH (30-41). It also shows no cross-reactivity with CRH-related substances from other species, i.e. urotensin I (fish) and sauvagine (frog). The cleavage site antibody (781), recognizes the full length proCRH molecule in Western blotting and in liquid phase radioimmunoassay from transfected CHO-K1 cells expressing the full length pre-proCRH cDNA. Using immunofluorescence and immunoprecipitation techniques followed by SDS-PAGE and autoradiography, we confirm the presence of the intact CRH precursor molecule within the nucleus and the cytoplasm of stably transfected CHO-K1 cells expr...Continue Reading

References

Jun 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·W S SossinR H Scheller
Jan 1, 1988·Annual Review of Physiology·B A Eipper, R E Mains
May 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·C PatzeltD F Steiner
Jan 1, 1980·Annals of the New York Academy of Sciences·J T PottsA Rich
Jan 1, 1982·Annual Review of Physiology·K Docherty, D F Steiner
Jul 1, 1995·The Journal of Endocrinology·M G Castro, E Morrison
Apr 1, 1993·Journal of Neuroimmunology·R EkmanH O Sjögren

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