Heterogeneity of contractile proteins. Purification and characterization of two species of troponin T from rabbit fast skeletal muscle.
Two species of troponin T have been purified by ion-exchange chromatography from erector spinae, the major fast white muscle of the rabbit back, and from a pool of the fast hindlimb muscles gastrocnemius and plantaris. Designated Tn-T1f and Tn-T2f, they can be resolved by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, with apparent molecular weights of 37,500 and 37,000 respectively. Their amino acid compositions are similar and correlate well with that reported for troponin T from fast muscle (Pearlstone, J. R., Carpenter, M. R., and Smillie, L. B. (1977) J. Biol. Chem. 252, 971-977). Tn-T2f most likely corresponds to the previously studied troponin T; further characterization was undertaken to determine how the newly identified Tn-T1f differs from Tn-T2f. Phosphorylation of alkaline phosphatase-treated troponin demonstrated that Tn-T1f and Tn-T2f are not interconverted by a change in phosphorylation state. Comparison of the CNBr fragments of Tn-T1f and Tn-T2f by SDS-gel electrophoresis and reverse phase high-performance liquid chromatography revealed similar but not identical peptide patterns. The major difference occurs in the amino-terminal CNBr peptides corresponding to CB3. Since both Tn-T1f and Tn-T2f have blocked amino termini, the difference does not result from proteolysis at the amino terminus of one of the proteins. These observations indicate that the two species of troponin T do not result from a known post-translational modification, but rather from differences in the amino acid sequence, suggesting that they arise either from the expression of different genes or a single gene from which different mRNAs are transcribed.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D009132:Muscles / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis; D011817:Rabbits; D014336:Troponin / Q000302:isolation & purification*; D020107:Troponin T
D000596:Amino Acids; D010446:Peptide Fragments; D014336:Troponin; D020107:Troponin T; D003488:Cyanogen Bromide