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>PARTIAL CHARACTERIZATION OF A PUTATIVE 110kDa MYOSIN FROM THE GREEN ALGACHARA CORALLINABYIN VITROBINDING OF FLUORESCENT F‐ACTIN
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PARTIAL CHARACTERIZATION OF A PUTATIVE 110kDa MYOSIN FROM THE GREEN ALGACHARA CORALLINABYIN VITROBINDING OF FLUORESCENT F‐ACTIN
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机译：PARTIAL CHARACTERIZATION OF A PUTATIVE 110kDa MYOSIN FROM THE GREEN ALGACHARA CORALLINABYIN VITROBINDING OF FLUORESCENT F‐ACTIN
AbstractUsing the binding of heterologous, rhodamine phalloidin‐labelled F‐actinin vitro, two F‐actin binding proteins were identified in protein extracts from the green algaChara corallinaafter fractionation by anion exchange chromatography. The first protein, a putative myosin, released laterally bound F‐actin at ATP‐concentrations as low as 1μm; equivalent concentrations of ADP were not effective. Binding of F‐actin was inhibited by the sulfhydryl‐alkylating agent N‐ethylmaleimide (NEM). Binding of F‐actin was also abolished by a monoclonal anti‐myosin (J14) previously used for immunodetection and immunolocalization in internodal cells (Groliget al., 1988,Eur J Cell Biol47: 22–31). Immunoblotting with J14 detected a 110kDa polypeptide only in those protein fractions that had revealed ATP‐sensitive binding of F‐actin. The putative myosin bound with mediocre affinity to immobilized calmodulin and free Ca2+‐concentration made no difference to this binding affinity. In contrast to the putative myosin, the second, less abundant protein revealed ATP‐insensitive and end‐wise binding to the microfilament and was not recogn
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