title: Production, crystallization, and preliminary X-ray analysis of rabbit skeletal muscle troponin complex consisting of troponin C and fragment (1-47) of troponin I.
abstract: Troponin is a ternary protein complex consisting of subunits TnC. TnI, and TnT, and plays a key role in calcium regulation of the skeletal and cardiac muscle contraction. In the present study, a partial complex (CI47) was prepared from Escherichia coli-expressed rabbit skeletal muscle TnC and fragment 1-47 of TnI, which is obtained by chemical cleavage of an E. coli-expressed mutant of rabbit skeletal muscle TnI. Within the ternary troponin complex, CI47 is thought to form a core that is resistant to proteolytic digestion, and the interaction within CI47 likely maintains the integrity of the troponin complex. Complex CI47 was crystallized in the presence of sodium citrate. The addition of trehalose improved the diffraction pattern of the crystals substantially. The crystal lattice belongs to the space group P3(1)(2)21, with unit cell dimensions a = b = 48.2 A, c = 162 A. The asymmetric unit presumably contains one CI47 complex. Soaking with p-chloromercuribenzenesulfonate (PCMBS) resulted in loss of isomorphism, but enhanced the quality of the crystals. The crystals diffracted up to 2.3 A resolution, with completeness of 91% and R(merge) = 6.4%. The crystals of PCMBS-derivative should be suitable for X-ray studies using the multiple-wavelength anomalous diffraction technique. This is the first step for elucidating the structure of the full troponin complex.
authors.affiliation: International Institute for Advanced Research, Matsushita Electric Industrial, Kyoto, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Saijo
authors.forename: S
authors.initials: S
authors.lastname: Takeda
authors.forename: A
authors.initials: A
authors.lastname: Scherer
authors.forename: T
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Maéda
authors.forename: H
authors.initials: H
authors.lastname: Taniguchi
authors.forename: M
authors.initials: M
authors.lastname: Yao
authors.forename: S
authors.initials: S
authors.lastname: Wakatsuki
mesh_terms: D000818:Animals; D018360:Crystallography, X-Ray; D018482:Muscle, Skeletal / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D011817:Rabbits; D019209:Troponin C / Q000737:chemistry*; D019210:Troponin I / Q000737:chemistry*
chemical_list: D010446:Peptide Fragments; D019209:Troponin C; D019210:Troponin I