Tropomyosin (TM) has been isolated from the cardiac muscle, and fast and slow trunk (myotomal) muscles of the mature salmonid fish Atlantic salmon (SuZmo suZur) and rainbow trout (Sulmo guirdneri). When examined electrophoretically, isoforms of TM were detected which were specific, and exclusive, to each type of muscle. Cardiac and fast muscles contained single and distinct isoforms, while slow muscle contained two distinct isoforms, closely related in terms of apparent M, and PI. There was no detectable difference between the same TM type from either salmon or trout. On a variety of gel systems, the cardiac and slow isoforms migrated in close proximity to each other and to rabbit a-TM. The fast isoform comigrated with rabbit /3-TM. In developing salmon fry, a more acidic (unphosphorylated) variant of TM was present in addition to, and of similar M , to, the fast adult isoform. This TM declined in steady-state level during maturation and was virtually undetected in adult muscle. All of the isolated TMs contained little or no covalently bound phosphate and were blocked at the N-terminus. The amino acids released by carboxypeptidase A, when ordered to give maximal similarity to other muscle TMs, were consistent with the following sequences: fast (LDNALNDMTSI) and cardiac (LDHALNDMTSL). The C-terminal region of the slow TM contained His but was heterogeneous. In viscosity measurements, performed as a function of increasing protein concentration, at low ionic strength (t = 5"C, pH 7.00), fast TM exhibited the highest relative viscosity values. Lower and equivalent levels of polymerisation occurred with the cardiac and slow TMs. Polymerisation of all three isoforms was temperature-dependent, with cardiac TM being least sensitive and fast TM being most sensitive. Determination of the complete coding sequence of adult fast TM confirmed the findings of the carboxypeptidase analysis, but the remainder of the sequence more closely resembled a-type TMs than /3-type TMs. Overall, salmon fast TM contains 20 (mostly conservative) substitutions compared to rabbit striated muscle a-TM and 40 (mostly conservative) substitutions compared to rabbit striated muscle /3-TM. This demonstrates that electrophoretic mobility is not, in all instances, a suitable method to assess the isomorphic nature of striated muscle TMs.

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Five major troponin-T isoforms were isolated from the myotomal muscles of Atlantic salmon: three from fast muscle (Tn-T1F, Tn-T2F and Tn-T3F) and two from slow muscle (Tn-T1S and Tn-T2S). In addition to their presence in troponin preparations, these proteins were also recognised to be Tn-T on the basis of immunoreaction with anti-troponin-T antibodies and partial amino acid sequence. The electrophoretic mobility in the presence of SDS of the various Tn-Ts increases in the order: 1S < 1F < 2S < 2F < or = 3F. Compositional analysis shows that the higher M(r) forms (1F and 1S) contain considerably more proline, glutamic acid and alanine than the lower-M(r) forms (2F, 3F and 2S). Every isoform lacks cysteine and phosphoserine is present only in isoforms 2F and 3F. All of the Tn-Ts, with the exception of isoform 1F, are N-terminally blocked. CNBr fragments from same cell type Tn-Ts yield identical sequences over at least fifteen Edman cycles. Two full-length cDNA sequences, presumed to represent 1S and 3F, or isoforms that are highly similar, are reported. As documented for higher vertebrate Tn-Ts, the predicted primary structures display a non-uniform distribution of charged amino acids and greater divergence at each end than in the central section. The most striking difference between the two salmonid proteins is the presence of a N-terminal (proline-, glutamic acid- and alanine-rich) extension of about fifty amino acids in Tn-T1s (278 amino acids) that is missing from the fast muscle Tn-T (223 amino acids). The sequences also differ in that 1S lacks the known phosphorylation site while the fast-type isoform contains serine next to the initiating methionine. Of the two, the slow isoform has accumulated the greater number of substitutions.