TY - JOUR
T1 - Synthetic peptide analogs of skeletal troponin C
T2 - Fluorescence studies of analogs of the low-affinity calcium-binding site II
AU - Kanellis, Panos
AU - Yang, Jessie
AU - Cheung, Herbert C.
AU - Lenkinski, Robert E.
PY - 1983/2/1
Y1 - 1983/2/1
N2 - Two 12-residue peptides were synthesized by the solid-phase method as structural analogs of a Ca2+-binding loop of rabbit skeletal troponin C. The sequence of the analogs corresponds to the binding loop of the Ca2+-specific low affinity binding site II (residues 63-74) but with two amino acid substitutions. In one analog, Phe-72 was replaced by tyrosine. In the other Gly-66 was substituted by serine and Phe-72 by tyrosine. The intrinsic fluorescence of the peptides was enhanced upon addition of Tb3+ or large excess of Ca2+. From the enhancement of Tb3+ emission association constants in the range (2-3) × 105m-1 and a binding stoichiometry of 1 were determined for Tb3+ binding to the peptides. Large excess of Ca2+ displaced Tb3+ from the Tb3+-peptide complexes and from these results apparent stability constants of 500-700 m-1 were deduced for Ca2+ binding. Preliminary proton nuclear magnetic resonance results on one of the peptides indicated that La3+ induced considerable perturbation of the amide proton resonances of several residues, including the aspartate at position 3, the tyrosine at position 10, and the two glutamates at the C-terminus. The results suggest involvement of these residues in cation coordination.
AB - Two 12-residue peptides were synthesized by the solid-phase method as structural analogs of a Ca2+-binding loop of rabbit skeletal troponin C. The sequence of the analogs corresponds to the binding loop of the Ca2+-specific low affinity binding site II (residues 63-74) but with two amino acid substitutions. In one analog, Phe-72 was replaced by tyrosine. In the other Gly-66 was substituted by serine and Phe-72 by tyrosine. The intrinsic fluorescence of the peptides was enhanced upon addition of Tb3+ or large excess of Ca2+. From the enhancement of Tb3+ emission association constants in the range (2-3) × 105m-1 and a binding stoichiometry of 1 were determined for Tb3+ binding to the peptides. Large excess of Ca2+ displaced Tb3+ from the Tb3+-peptide complexes and from these results apparent stability constants of 500-700 m-1 were deduced for Ca2+ binding. Preliminary proton nuclear magnetic resonance results on one of the peptides indicated that La3+ induced considerable perturbation of the amide proton resonances of several residues, including the aspartate at position 3, the tyrosine at position 10, and the two glutamates at the C-terminus. The results suggest involvement of these residues in cation coordination.
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U2 - 10.1016/0003-9861(83)90444-7
DO - 10.1016/0003-9861(83)90444-7
M3 - Article
C2 - 6824337
AN - SCOPUS:0020595889
VL - 220
SP - 530
EP - 540
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
SN - 0003-9861
IS - 2
ER -