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NMR identification of calcineurin B residues affected by binding of a calcineurin A peptide
Author(s) -
Anglister Jacob,
Ren Hao,
Klee Claude B.,
Bax Ad
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01192-h
Subject(s) - calcineurin , peptide , chemistry , chaps , nuclear magnetic resonance spectroscopy , biochemistry , stereochemistry , membrane , transplantation , medicine , surgery
Triple resonance 3D NMR methods have been used to study the interaction between calcineurin B and a peptide fragment of calcineurin A for which it has high affinity ( K D ∼4 × 10 −7 M). Although calcineurin B aggregates at NMR concentrations of ∼ 1 mM, in the presence of a target peptide fragment of calcineurin A it becomes monomeric and yields NMR spectra that are very similar to those reported previously for calcineurin B solubilized by the zwitterionic detergent CHAPS. Changes in chemical shifts between CHAPS‐ and peptide‐solubilized calcineurin B are small which is indicative of no differences in secondary structure. Residues most affected by binding to target peptide are found primarily on the hydrophobic faces of the four helices, present in each of the two globular domains in calcineurin B, and in the loops connecting helices II and III, IV and V, and possibly in the C‐terminal 12 residues, which also exhibit a change in mobility.