Processing of the atrial natriuretic factor propeptide by atrial cardiocytes as revealed by immunocryoultramicrotomy.
Antibodies were raised against three fragments of the rat ANF molecule: C-terminal atrial natriuretic factor (ANF)-(101-126), N-terminal ANF-(11-37), and the putative cleavage site of the ANF propeptide, ANF-(94-103). These antibodies were purified by affinity chromatography and revealed a major band (17K) corresponding to the propeptide by Western blot analysis. Antibodies against ANF-(94-103) were used in a RIA. All of the peptides that possess this region (98-99) were able to displace iodinated pro-ANF from the antibody. However, peptides that have only one part of the fragment, such as ANF-(99-126) or ANF-(1-98), failed to displace pro-ANF; human ANF-(79-98) (100 pmol) demonstrated about 0.01% cross-reactivity. Immunohistochemical studies revealed that all atrial cardiocytes are reactive with the three antibodies. Immunocryoultramicrotomy revealed that the propeptide travels, uncleaved, from the Golgi complex to immature granules and mature secretory granules. These results indicate that cleavage of the ANF propeptide does not occur at these sites.
Thibault
G
G
Laboratory of Pathobiology, Clinical Research Institute of Montreal, Quebec, Canada.
Haile-Meskel
H
H
Wrobel-Konrad
E
E
Ballak
M
M
Garcia
R
R
Genest
J
J
Cantin
M
M
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D009320:Atrial Natriuretic Factor / Q000032:analysis / Q000235:genetics*; D003429:Cross Reactions; D005260:Female; D006325:Heart Atria / Q000378:metabolism; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D009206:Myocardium / Q000378:metabolism* / Q000648:ultrastructure; D011498:Protein Precursors / Q000032:analysis / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains
D000906:Antibodies; D011498:Protein Precursors; D009320:Atrial Natriuretic Factor