Skeletal structure of clathrin triskelion in solution: experimental and theoretical approaches.
The physicochemical properties of the clathrin triskelion were determined by dynamic and static light-scattering and sedimentation analyses in Tris and triethanolamine (TEA) buffers of about pH 8, in which the clathrin triskelion has been found to be in different conformational states by electron microscopy [Heuser, J., & Kirchhausen, T. (1985) J. Ultrastruct. Res. 92, 1-27]. Dynamic light-scattering measurements provided diffusion coefficients (D0(20,w)) of 1.22 x 10(-7) and 1.23 x 10(-7) cm2/s, and ultracentrifugal analysis gave sedimentation coefficients (S0(20,w)) of 8.39 and 8.32 S in Tris and TEA buffer, respectively. The average Stokes radius of the protein was determined to be 175 A from its diffusion and sedimentation coefficients and its molecular weight. Static light-scattering analysis provided molecular weights of 6.58 x 10(5) and 6.41 x 10(5) and radii of gyration of 311 and 301 A in the respective buffers. These results indicate that the clathrin triskelion has a similar conformation in the two buffers. For clarification of the skeletal structure of the clathrin triskelion in solution, the physicochemical parameters were calculated by using two models in which the clathrin arms are bent at various angles in a plane, on the basis of the Bloomfield approximation and a formula derived to estimate the radius of gyration of proteins consisting of various structural units. Values for the Stokes radius, diffusion and sedimentation coefficients, and radius of gyration in the ranges of 178-170 A, (1.20-1.26) x 10(-7) cm2/s, 8.26-8.66 S, and 316-266 A, respectively, were obtained with these models with the arms bent in the range of 0-60 degrees.(ABSTRACT TRUNCATED AT 250 WORDS)
Yoshimura
T
T
Institute for Enzyme Research, University of Tokushima, Japan.
Kameyama
K
K
Maezawa
S
S
Takagi
T
T
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002966:Clathrin / Q000737:chemistry*; D004058:Diffusion; D004983:Ethanolamines / Q000737:chemistry; D008958:Models, Molecular; D011487:Protein Conformation; D012996:Solutions; D013329:Structure-Activity Relationship; D014325:Tromethamine; D014461:Ultracentrifugation
D002966:Clathrin; D004983:Ethanolamines; D012996:Solutions; D014325:Tromethamine; C009546:triethanolamine