Structure, binding interface and hydrophobic transitions of Ca2+-loaded calbindin-D(28K).
Calbindin-D(28K) is a Ca2+-binding protein, performing roles as both a calcium buffer and calcium sensor. The NMR solution structure of Ca2+-loaded calbindin-D(28K) reveals a single, globular fold consisting of six distinct EF-hand subdomains, which coordinate Ca2+ in loops on EF1, EF3, EF4 and EF5. Target peptides from Ran-binding protein M and myo-inositol monophosphatase, along with a new target from procaspase-3, are shown to interact with the protein on a surface comprised of alpha5 (EF3), alpha8 (EF4) and the EF2-EF3 and EF4-EF5 loops. Fluorescence experiments reveal that calbindin-D(28K) adopts discrete hydrophobic states as it binds Ca2+. The structure, binding interface and hydrophobic characteristics of Ca2+-loaded calbindin-D(28K) provide the first detailed insights into how this essential protein may function. This structure is one of the largest high-resolution NMR structures and the largest monomeric EF-hand protein to be solved to date.
Kojetin
Douglas J
DJ
Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, North Carolina 27695, USA.
Venters
Ronald A
RA
Kordys
David R
DR
Thompson
Richele J
RJ
Kumar
Rajiv
R
Cavanagh
John
J
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D064026:Calbindins; D002118:Calcium / Q000378:metabolism*; D003001:Cloning, Molecular; D006860:Hydrogen Bonding; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry; D017510:Protein Folding; D017433:Protein Structure, Secondary; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000737:chemistry* / Q000378:metabolism*
D064026:Calbindins; D010455:Peptides; D011994:Recombinant Proteins; D064030:S100 Calcium Binding Protein G; D002118:Calcium