Protein p67. A calcium-binding protein localized at the sarcolemma of secretory atrial myocytes.
Bovine heart 67-kd protein (p67) was coisolated with calpactin I complex by cycles of Ca(2+)-dependent precipitation followed by solubilization with EGTA-containing buffer. Using affinity-purified anti-p67 antibody and anti-p36 (36-kd subunit of calpactin I) antibody, we examined the localization of the two proteins in secretory atrial myocytes and other endocrine tissues of adult rats. Immunofluorescence microscopy showed that p67 was expressed both in the atrial myocytes in situ and in cultured atrial myocytes in which we failed to detect p36 and that p67 appeared to be closely associated with the cell surface. We also found that p67 was colocalized with p36 in the thyroid follicle epithelium and zona reticularis of the adrenal gland. On the other hand, neither p67 nor p36 was detectable in pancreas islet cells. Immunoelectron microscopy revealed that p67 was localized at the sarcolemma in the atrial myocytes in situ. The p67, which was shown to be a globular molecule with a diameter of 18-25 nm by a low-angle rotary shadowing method, bound radioactive Ca2+ on a nitrocellulose membrane. The results suggest that Ca(2+)-binding proteins expressed in endocrine cells seem to vary from tissue to tissue and that p67 may function in Ca(2+)-mediated events at the plasma membrane of secretory atrial myocytes and some types of endocrine cells expressing this protein.
Iida
H
H
Department of Anatomy, Faculty of Medicine, Kyushu University, Japan.
Nishitani
H
H
Shibata
Y
Y
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D017302:Annexins; D000906:Antibodies / Q000276:immunology; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D002478:Cells, Cultured; D004702:Endocrine Glands / Q000378:metabolism; D006325:Heart Atria; D008970:Molecular Weight; D009206:Myocardium / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D012508:Sarcolemma / Q000378:metabolism*
D000596:Amino Acids; D017302:Annexins; D000906:Antibodies; D002135:Calcium-Binding Proteins