title: Orientation of actin monomer in the F-actin filament: radial coordinate of glutamine-41 and effect of myosin subfragment 1 binding on the monomer orientation.
abstract: We have employed the method of radial distance measurements in order to orient the actin monomer in the F-actin filament. This method utilizes fluorescence resonance energy transfer measurements of the distance between two equivalent chemical points located on two different monomers. The interprobe distance obtained this way is used to compute the radial coordinate of the labeled amino acid [Taylor, D. L., Reidler, J., Spudich, J. A., & Stryer, L. (1981) J. Cell Biol. 89, 362-367]. Theoretical analysis has indicated that if radial coordinates of four points are determined and six intramolecular distances are known, one can, within symmetry limits, position the monomer about the filament axis. The radial distance of Gln-41 that had been enzymatically modified with dansyl, rhodamine, and fluorescein derivatives of cadaverine was found to be approximately 40-42 A. The determination of the radial distance of Cys-374 was accomplished by using monobromobimane and N-[[(iodoacetyl)amino]ethyl]-5- naphthylamine-1-sulfonate as donors and N-[4-[[4-(dimethylamino)phenyl]azo]phenyl]maleimide as acceptor; the results were consistent with a radial coordinate for this residue of 20-25 A. The effect of myosin subfragment 1 (S1) binding on the radial coordinates of (1) Gln-41, (2) Cys-374, and (3) the nucleotide binding site was also examined. S1 had a small effect on the radial coordinate of Gln-41, increasing it to 44-47 A. In the two remaining lases the change in the radial coordinate due to the S1 binding was negligible. This finding excludes certain models of the interaction between actin and S1 in which actin monomer rotates by a large angle when subfragment 1 binds to it.
authors.affiliation: Cardiovascular Research Institute, University of California, San Francisco 94143-0524.
authors.forename: A A
authors.initials: AA
authors.lastname: Kasprzak
authors.forename: R
authors.initials: R
authors.lastname: Takashi
authors.forename: M F
authors.initials: MF
authors.lastname: Morales
mesh_terms: D000199:Actins; D000244:Adenosine Diphosphate / Q000378:metabolism; D001667:Binding, Competitive; D003545:Cysteine; D004735:Energy Transfer; D005973:Glutamine; D015879:Myosin Subfragments; D009218:Myosins / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D000199:Actins; D015879:Myosin Subfragments; D009711:Nucleotides; D010446:Peptide Fragments; D005973:Glutamine; D000244:Adenosine Diphosphate; D009218:Myosins; D003545:Cysteine