NMR identification of calcineurin B residues affected by binding of a calcineurin A peptide

FEBS Letters

Bax, Ad¹  Anglister, Jacob¹  Klee, Claude B.²  Ren, Hao² 

[1] Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0520, USA;Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-0520, USA

关键词: Calcineurin;    NMR;    CHAPS;    Peptide-protein interaction;    CBCA(CO)NH;    3D (13Cα;    13Cβ)-15N-1H spectrum correlating amide 1H and 15N with Cα and Cβ shifts of the preceding residue;    CBCANH;    3D (13Cα;    13Cβ)-15N-1H spectrum correlating amide 1H and 15N with sequential and intraresidue Cα and Cβ shifts;    CnA;    calcineurin A;    CnB;    calcineurin B;    CHAPS;    3-[(3-cholamidopropyl)-dimethyl-ammonio]-1-propanesulfonate;    FKBP;    FK506-binding protein;    HNCA;    3D 1H-15N-13Cα correlation;    HSQC;    heteronuclear single quantum correlation;    NF-AT;    nerve factor of activated T cells;    NF-ATc;    cytoplasmic subunit of NF-AT;   

DOI  :  10.1016/0014-5793(95)01192-H

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown