Antibodies raised against a peptide corresponding to a Gs alpha domain reveal a vimentin-associated protein.
In an attempt to localize the guanine nucleotide binding protein Gs alpha by immunocytochemistry, we synthetized peptides corresponding to several stretches of residues deduced from the published cDNA sequence of Gs alpha and raised antibodies against them. Among the peptides, one corresponding to residues 367-381 elicited antibodies that immunocytochemically detected, at the optical level, what appeared to be vimentin in several cells and tissues. Studies at the ultrastructural level confirmed this observation and also showed weak staining of some areas of plasma membranes of glial and nerve cells. Analysis by Western blots of rat brain subcellular fractions indicated that: (a) the protein stained by the anti-peptide antibodies was associated with the cytoskeleton; and (b) it was not vimentin but a protein of higher molecular weight, 65 KD. We accordingly suggest that the Gs alpha-derived peptide elicited two types of antibodies, one recognizing Gs alpha in fixed tissues, the other recognizing an epitope located in a vimentin-associated protein. This study emphasizes the caution that is needed when conclusions are drawn on the basis of immunocytochemical studies using antipeptide antibodies.
Anglade
G
G
Laboratoire de Biochimie, URA, CNRS 1455, Faculté de Médecine Secteur Nord, Marseille, France.
Dupouey
P
P
Ensergueix
D
D
Ceard
B
B
Monneron
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D001923:Brain Chemistry; D002531:Cerebellum / Q000737:chemistry; D003599:Cytoskeleton / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D019204:GTP-Binding Proteins / Q000032:analysis*; D000882:Haplorhini; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D051381:Rats; D017208:Rats, Wistar; D014746:Vimentin / Q000032:analysis*
D000911:Antibodies, Monoclonal; D014746:Vimentin; D019204:GTP-Binding Proteins