Characterization of chicken gizzard calcyclin and examination of its interaction with caldesmon.
Using a procedure developed to purify calcyclin from mouse Ehrlich ascites tumor cells calcyclin was purified from smooth muscle of chicken gizzard. Chicken gizzard calcyclin bound to phenyl-Sepharose in a calcium dependent manner as did mouse EAT cells and rabbit lung calcyclin but appeared to be more acidic than its mammalian counterparts as revealed by ion exchange chromatography on Mono Q. Chicken gizzard calcyclin bound 45Ca2+ on nitrocellulose filters and exhibited a shift in electrophoretic mobility on urea-PAGE depending on Ca2+ concentration. Crosslinking experiments with BS3 showed that chicken gizzard calcyclin was able to form noncovalent dimers. As indicated by a decrease in maximum tryptophan fluorescence emission of caldesmon (about 14% at 1:1 molar ratio) and displacement of calmodulin from its complex with caldesmon, chicken gizzard calcyclin binds caldesmon. This binding was, however, much weaker than that of calmodulin and could not influence the interaction of caldesmon with actin. In consequence, calcyclin was unable to reverse the inhibitory effect of caldesmon on actin-activated Mg(2+)-ATPase activity of myosin in the presence of Ca2+.
Filipek
A
A
Department of Muscle Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland.
Zasada
A
A
Wojda
U
U
Makuch
R
R
Dabrowska
R
R
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism*; D018797:Cell Cycle Proteins; D002645:Chickens / Q000378:metabolism*; D004789:Enzyme Activation; D005895:Gizzard, Avian / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009218:Myosins / Q000378:metabolism*; D011817:Rabbits; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; D013050:Spectrometry, Fluorescence; D014335:Tropomyosin / Q000378:metabolism
D000199:Actins; D002135:Calcium-Binding Proteins; D002148:Calmodulin-Binding Proteins; D018797:Cell Cycle Proteins; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C489133:S100A6 protein, Gallus gallus; C489134:S100a6 protein, mouse; D014335:Tropomyosin; D009218:Myosins; D002118:Calcium