Calcium binding mode of gamma-carboxyglutamic acids in conantokins.
Conantokin-T (con-T) and conantokin-G (con-G) are two highly homologous peptide toxins found in Conus venom. The former is a 21-residue peptide with four gamma-carboxyglutamic acid (Gla) residues (at positions 3, 4, 10 and 14), while the latter is a 17-residue peptide with five gamma-carboxyglutamic acid residues (at positions 3, 4, 7, 10 and 14). Despite the apparent similarity in number and relative positions of the gamma-carboxyglutamic acid residues, (113)Cd-NMR studies indicated a distinct metal binding behavior for con-G and con-T. There appears to be four binding sites in con-G in contrast to one metal binding site in con-T. To elucidate the mode of calcium binding by the gamma-carboxyglutamic acid residues in these conantokins, we designed various analogous peptides with their gamma-carboxyglutamic acid replaced by other amino acid residues. (113)Cd-NMR experiments on conantokin analogues reveal that the major difference in the number of metal binding sites between con-G and con-T is due to the residue at position 7. We also performed molecular simulations to calculate the relative binding free energies of several potential binding sites. Based on our theoretical and experimental results, we propose a 'four-site' binding model for conantokin-G and a 'single-site' binding model for conantokin-T.
Lin
C H
CH
Department of Life Sciences, National Tsing-Hua University,Hsin-Chu 30043, Taiwan.
Chan
F C
FC
Hwang
J K
JK
Lyu
P C
PC
D015055:1-Carboxyglutamic Acid / Q000378:metabolism*; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001665:Binding Sites; D002104:Cadmium; D002118:Calcium / Q000378:metabolism*; D002942:Circular Dichroism; D003198:Computer Simulation; D020916:Conotoxins; D018698:Glutamic Acid; D036341:Intercellular Signaling Peptides and Proteins; D007554:Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008978:Mollusk Venoms / Q000737:chemistry* / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation
D020916:Conotoxins; D036341:Intercellular Signaling Peptides and Proteins; D007554:Isotopes; D008978:Mollusk Venoms; D010455:Peptides; D010456:Peptides, Cyclic; D002104:Cadmium; C063571:conantokin-T; D018698:Glutamic Acid; D015055:1-Carboxyglutamic Acid; C055407:conotoxin GV; D002118:Calcium