The (1)H NMR structure of bovine Pb(2+)-osteocalcin and implications for lead toxicity.
Structural information on the effect of Pb(2+) on proteins under physiologically relevant conditions is largely unknown. We have previously shown that low levels of lead increased the amount of osteocalcin bound to hydroxyapatite (BBA 1535:153). This suggested that lead induced a more compact structure in the protein. We have determined the 3D structure of Pb(2+)-osteocalcin (49 amino acids), a bone protein from a target tissue, using (1)H 2D NMR techniques. Lead, at a stoichiometry of only 1:1, induced a similar fold in the protein as that induced by Ca(2+) at a stoichiometry of 3:1. The structure consisted of an unstructured N-terminus and an ordered C-terminal consisting of a hydrophobic core (residues 16-49). The genetic algorithm-molecular dynamics simulation predicted the lead ion was coordinated by the Gla 24 and Gla 21 residues. It is proposed that mineral binding occurs via uncoordinated Gla oxygen ions binding to calcium in hydroxyapatite. A comparison of Pb(2+)- and Ca(2+)-osteocalcin suggests Pb(2+), at a lower stoichiometry, may induce similar conformational changes in proteins and subsequent molecular processes normally controlled by calcium alone. This may contribute to a molecular mechanism of lead toxicity for calcium binding proteins. Lead exposure may alter the amount of mineral bound osteocalcin and contribute to abnormal bone remodeling.
Dowd
T L
TL
Department of Chemistry Room 359 NE, Brooklyn College of the City University of New York, 2900 Bedford Avenue, Brooklyn, New York 11210, USA. TDowd@brooklyn.cuny.edu
Li
L
L
Gundberg
C M
CM
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D006859:Hydrogen / Q000737:chemistry; D007854:Lead / Q000737:chemistry / Q000378:metabolism*; D007855:Lead Poisoning / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D015675:Osteocalcin / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding
D015675:Osteocalcin; D007854:Lead; D006859:Hydrogen; D002118:Calcium
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