Characterization of the single tyrosine containing troponin C from lungfish white muscle. Comparison with several fast skeletal muscle troponin C's from fish species.
Troponin C molecules from fast skeletal muscle of the following fish species (trout, whiting, lungfish, tilapia, and cod) have been purified to homogeneity. Upon binding of Ca2+ or Mg2+, lungfish troponin C is the only troponin C from fish white muscle to show the typical increase of tyrosine fluorescence emission quantum yield reported for rabbit fast skeletal muscle troponin C. The increase of tyrosine fluorescence signal occurring upon Ca2+ and Mg2+ titration of lungfish troponin C has been used to determine the corresponding affinity constants. With K(Ca) = 7.0 10(7) M-1 and K(Mg) = 3.6 10(3) M-1, the sites probed by the tyrosine residue of lungfish troponin C are typical of the COOH-terminal domain of fast skeletal troponin C's. The amino acid sequencing of the tyrosine containing tryptic peptides has allowed us to position the single tyrosine residue at position 7 in the Ca2+ binding loop of the third site, in identical position to Tyr109 of troponin C from rabbit fast skeletal muscle. Metal ion binding studies followed by intrinsic fluorescence or Tb3+ luminescence indicate that the conformation of the structural domain of lungfish troponin C with one metal ion bound is close to the physiological conformation of this domain.
François
J M
JM
Laboratoire de Biochimie, Université de Liège, Belgium. Jean-Marie.Francois@ulg.ac.be
Altintas
A
A
Gerday
C
C
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000378:metabolism; D005399:Fishes / Q000502:physiology*; D008274:Magnesium / Q000378:metabolism; D008969:Molecular Sequence Data; D018656:Muscle Fibers, Fast-Twitch / Q000737:chemistry*; D018482:Muscle, Skeletal / Q000737:chemistry*; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D013057:Spectrum Analysis / Q000379:methods; D013725:Terbium / Q000378:metabolism; D019209:Troponin C / Q000737:chemistry* / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism
D000596:Amino Acids; D019209:Troponin C; D013725:Terbium; D014443:Tyrosine; D008274:Magnesium; D002118:Calcium