Characterization and confocal imaging of calponin in gastrointestinal smooth muscle.
Calponin isolated from chicken gizzard smooth muscle binds in vitro to actin in a Ca(2+)-independent manner and thereby inhibits the actin-activated Mg(2+)-adenosinetriphosphatase of smooth muscle myosin. This inhibition is relieved when calponin is phosphorylated by protein kinase C or Ca2+/calmodulin-dependent protein kinase II, suggesting that calponin is involved in thin filament-associated regulation of smooth muscle contraction. To further examine this possibility, calponin was isolated from toad stomach smooth muscle, characterized biochemically, and localized in intact isolated cells. Toad stomach calponin had the same basic biochemical properties as calponin from other sources. Confocal immunofluorescence microscopy revealed that calponin in intact smooth muscle cells was localized to long filamentous structures that were colabeled by antibodies to actin or tropomyosin. Preservation of the basic biochemical properties of calponin from species to species suggests that these properties are relevant for its in vivo function. Its colocalization with actin and tropomyosin indicates that calponin is associated with the thin filament in intact smooth muscle cells.
Walsh
M P
MP
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
Carmichael
J D
JD
Kargacin
G J
GJ
D000199:Actins / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002025:Bufonidae; D002135:Calcium-Binding Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D005753:Gastric Mucosa / Q000378:metabolism*; D005895:Gizzard, Avian / Q000378:metabolism; D007150:Immunohistochemistry; D008840:Microfilament Proteins; D009124:Muscle Proteins / Q000378:metabolism*; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D013270:Stomach / Q000166:cytology; D000096985:Calponins
D000199:Actins; D000906:Antibodies; D002135:Calcium-Binding Proteins; D008840:Microfilament Proteins; D009124:Muscle Proteins; D011493:Protein Kinase C; D000096985:Calponins