Nuclear magnetic resonance studies of the structure of B50/neuromodulin and its interaction with calmodulin.
B50/neuromodulin is a neuronal phosphoprotein that is found in association with the inner membrane of nerve cells. In this work, we have studied the structure of bovine B50 in aqueous solution (pH 7.5) by 1H nuclear magnetic resonance (NMR) spectroscopy and our results indicate that B50 is an unstructured protein under these conditions. One-dimensional 1H-NMR titration studies of the interaction between B50 and calmodulin (CaM) have shown that B50 does not interact with (or) interacts very weakly with apo-CaM in solution; neither does B50 interact with Ca(2+)-CaM. These NMR data are consistent with an earlier observation that B50 is not capable of binding apo-CaM in vitro unless some nonionic detergent is present. We have also detected aromatic NMR peaks for a new posttranslational modification that might involve the His residues of the protein. The interaction of a 14-residue peptide (I38-L51) encompassing the CaM-binding domain of B50 with CaM was also studied by NMR. We have found from two-dimensional transferred nuclear Overhauser enhancement experiments that the B50 peptide binds weakly to apo-CaM in an alpha-helical conformation; the alpha-helix appears to be induced by the binding of the peptide to apo-CaM.
Zhang
M
M
Department of Biological Sciences, University of Calgary, Canada.
Vogel
H J
HJ
Zwiers
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002147:Calmodulin / Q000378:metabolism*; D002417:Cattle; D002942:Circular Dichroism; D003903:Deuterium; D019922:GAP-43 Protein; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism*; D017433:Protein Structure, Secondary; D012996:Solutions; D014867:Water
D002147:Calmodulin; D019922:GAP-43 Protein; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D012996:Solutions; D014867:Water; D006639:Histidine; D003903:Deuterium