Characterization of the calcyclin (S100A6) binding site of annexin XI‐A by site‐directed mutagenesis
Open Access
- 5 February 1999
- journal article
- Published by Wiley in FEBS Letters
- Vol. 444 (1) , 11-14
- https://doi.org/10.1016/s0014-5793(99)00014-9
Abstract
Residues in annexin XI‐A essential for binding of calcyclin (S100A6) were examined by site‐directed mutagenesis. GST fusion proteins with the calcyclin binding site of annexin XI‐A, GST‐AXI 34–62 and GST‐AXI 49–77 bound to calcyclin‐Sepharose Ca2+‐dependently. The mutants GST‐AXI L52E, M55E, A56E and M59E lost the binding ability, whereas GST‐AXI A57E retained the ability. These results demonstrate that the hydrophobic residues L52, M55, A56 and M59 on one side surface of the α‐helix are critical for the binding. Assays with GST fusion proteins and synthesized peptides corresponding to the calcyclin binding site indicated that other regions around the calcyclin binding site are important to stabilize the conformation.Keywords
This publication has 36 references indexed in Scilit:
- An interaction‐based analysis of calcium‐induced conformational changes in Ca2+ sensor proteinsProtein Science, 1998
- The Role of Annexin II Tetramer in the Activation of PlasminogenJournal of Biological Chemistry, 1998
- 1H NMR assignments of apo calcyclin and comparative structural analysis with calbindin D9k and S100βProtein Science, 1996
- Annexin II in exocytosis: catecholamine secretion requires the translocation of p36 to the subplasmalemmal region in chromaffin cells.The Journal of cell biology, 1996
- The S100 family of EF-hand calcium-binding proteins: functions and pathologyTrends in Biochemical Sciences, 1996
- The S100 family of EF-hand calcium-binding proteins: functions and pathologyTrends in Biochemical Sciences, 1996
- Calcium-dependent Binding of S100C to the N-terminal Domain of Annexin IPublished by Elsevier ,1996
- The structure of calcyclin reveals a novel homodimeric fold for S100 Ca2+-binding proteinsNature Structural & Molecular Biology, 1995
- Binding Site of Annexin XI on the Calcyclin MoleculeBiochemical and Biophysical Research Communications, 1993
- Specific binding of CAP-50 to calcyclinFEBS Letters, 1992