Polylysine activates smooth muscle myosin ATPase activity via induction of a 10S to 6S transition.
Polylysine (10-13 kDa) stimulates contraction in smooth muscle skinned fibers and activates actomyosin adenosinetriphosphatase (ATPase) activity in the absence of myosin light chain phosphorylation [P. T. Szymanski and R. J. Paul. Adv. Exp. Med. 304: 363-368, 1991; P. T. Szymanski, J. D. Strauss, G. Doerman, J. DiSalvo, and R. J. Paul. Am J. Physiol. 262 (Cell Physiol. 31): C1445-C1455, 1992]. To provide further information on the mechanism of polylysine action on contractility in smooth muscle, we investigated its effect on ATPase activity and conformation of purified gizzard myosin. We report here that polylysine directly stimulates myosin ATPase activity in a concentration-dependent manner. This stimulation could be completely abolished with the addition of heparin, a negatively charged heteropolysaccharide. Polylysine (10 microM) increases myosin ATPase activity to a level similar to that of myosin phosphorylation. Addition of 10 microM polylysine to phosphorylated myosin [with myosin light chain kinase and adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), to approximately 1.9 mol P/mol myosin], however, did not further stimulate ATPase activity. At 0.2 M KCl (the salt concentration at which myosin exists primary in the 10S form), the addition of polylysine increases myosin ATPase activity to a level comparable to that of untreated myosin in 0.3 M KCl. These changes parallel the increase in solution viscosity elicited by polylysine. These results suggest that polylysine induces a transition in myosin conformation from the 10S to the 6S form, and this was confirmed by electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)
Szymanski
P T
PT
Department of Physiology and Biophysics, College of Medicine, University of Cincinnati, Ohio 45267-0576.
Ferguson
D G
DG
Paul
R J
RJ
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002645:Chickens; D005895:Gizzard, Avian / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D008854:Microscopy, Electron; D008968:Molecular Conformation; D009130:Muscle, Smooth / Q000201:enzymology*; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D010766:Phosphorylation; D011107:Polylysine / Q000494:pharmacology*; D011189:Potassium Chloride / Q000494:pharmacology
D011107:Polylysine; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D006493:Heparin; D000251:Adenosine Triphosphatases; D009218:Myosins