Proton NMR sequential resonance assignments, secondary structure, and global fold in solution of the major (trans-Pro43) form of bovine calbindin D sub 9k

Journal Article · Tue Aug 22 04:00:00 EDT 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00443a043· OSTI ID:5299135

Koerdel, J; Forsen, S; Chazin, W J ^[1]

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1. Research Institute of Scripps Clinic, La Jolla, CA (USA)

A wide range of two-dimensional {sup 1}H NMR experiments have been used to completely assign the 500-MHz {sup 1}H NMR spectrum of recombinant Ca{sup 2+}-saturated bovine calbindin D{sub 9k}. In solution, calbindin D{sub 9k} exists as an equilibrium mixture of isoforms with trans (75%) and cis (25%) isomers of the peptide bond at Pro43 which results in two sets of {sup 1}H NMR signals from approximately half of the amino acids. The complete {sup 1}H NMR assignments for the major, trans-pro43 isoform are presented here. By use of an integrated strategy for spin system identification, 62 of the 76 spin systems could be assigned to the appropriate residue type. Sequence-specific assignments were then obtained by the standard method. Secondary structure elements were identified on the basis of networks of sequential and medium-range nuclear Overhauser effects (NOEs), {sup 3}J{sub HN{alpha}} spin coupling constants, and the location of slowly exchanging amide protons. Four helical segments and a short {beta}-sheet between the two calcium binding loops are found. These elements of secondary structure and a few additional long-range NOEs provide the global fold. Good agreement is found between the solution and crystal structures of the minor A form of bovine calbindin D{sub 9k} and between the solution structures of the minor A form of bovine calbindin D{sub 9k} and intact porcine calbindin D{sub 9k}.

• OSTI ID: 5299135
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 28:17; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
ALKALINE EARTH METAL COMPOUNDS
AMINO ACIDS
ANIMALS
BARYONS
CALCIUM COMPOUNDS
CARBOXYLIC ACIDS
CATIONS
CATTLE
CHARGED PARTICLES
CHEMICAL BONDS
CHEMICAL SHIFT
DOMESTIC ANIMALS
ELEMENTARY PARTICLES
FERMIONS
HADRONS
IONS
LEUCINE
MAGNETIC RESONANCE
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MOLECULAR STRUCTURE
NMR SPECTRA
NUCLEAR MAGNETIC RESONANCE
NUCLEONS
ORGANIC ACIDS
ORGANIC COMPOUNDS
OVERHAUSER EFFECT
PROTEINS
PROTONS
RESONANCE
RUMINANTS
SPECTRA
SWINE
VALINE
VERTEBRATES