title: Functional homodimers and heterodimers of recombinant smooth muscle tropomyosin.
abstract: Skeletal and smooth muscle tropomyosin (Tm) require acetylation of their N-termini to bind strongly to actin. Tm containing an N-terminal alanine-serine (AS) extension to mimic acetylation has been widely used to increase binding. The current study investigates the ability of an N-terminal AS extension to mimic native acetylation for both alpha alpha and beta beta smooth Tm homodimers. We show that (1) AS alpha-Tm binds actin 100-fold tighter than alpha-Tm and 2-fold tighter than native smooth alphabeta-Tm, (2) beta-Tm requires an AS extension to bind actin, and (3) AS beta-Tm binds actin 10-fold weaker than AS alpha-Tm. Tm is present in smooth muscle tissues as >95% heterodimer; therefore, we studied the binding of recombinant alphabeta heterodimers with different AS extensions. This study shows that recombinant Tm requires an AS extension on both alpha and beta chains to bind like native Tm and that the alpha chain contributes more to actin binding than the beta chain. Once assembled onto an actin filament, all smooth muscle Tm's regulate S1 binding to actin Tm in the same way, irrespective of the presence of an AS extension.
authors.affiliation: Department of Biosciences, University of Kent, Canterbury CT2 7NY, UK.
authors.forename: Arthur
authors.initials: A
authors.lastname: Coulton
authors.forename: Sherwin S
authors.initials: SS
authors.lastname: Lehrer
authors.forename: Michael A
authors.initials: MA
authors.lastname: Geeves
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D017931:DNA Primers; D019281:Dimerization; D005895:Gizzard, Avian; D007700:Kinetics; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014335:Tropomyosin / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000199:Actins; D017931:DNA Primers; D011994:Recombinant Proteins; D014335:Tropomyosin