Circular dichroism and H-1 nuclear magnetic resonance studies on the solution and membrane structures of GAP-43 calmodulin-binding domain

被引:32
作者
Hayashi, N [1 ]
Matsubara, M [1 ]
Titani, K [1 ]
Taniguchi, H [1 ]
机构
[1] FUJITA HLTH UNIV,SCH MED,DIV BIOMED POLYMER SCI,INST COMPREHENS MED SCI,TOYOAKE,AICHI 47011,JAPAN
关键词
D O I
10.1074/jbc.272.12.7639
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Growth associated protein-43 (GAP 43) is believed to be palmitoylated near the N terminus and the modification is assumed to be involved in the membrane anchoring of the protein, However, GAP-43 isolated from bovine brain is not palmitoylated as shown by mass spectrometric analysis, but still retains the ability to bind phospholipids, suggesting that other parts of the molecule are involved in the interaction, Upon addition of acidic phospholipids, purified GAP-43 showed a conformational change from random coil to alpha-helix as indicated by a change in CD spectra. A synthetic peptide corresponding to the calmodulin-binding domain showed a similar conformational change from random coil to alpha-helix in the presence of various acidic phospholipids, These results suggest that the calmodulin-binding domain of GAP 43 is directly involved in the GAP-43-membrane interaction and undergoes a conformational change upon binding to phospholipid membranes. After phosphorylation by protein kinase C, the phospholipid-induced conformational changes were no longer observed. Structural characteristics of the calmodulin-binding domain peptide in aqueous and hydrophobic solvents were further studied in detail by two-dimensional H-1 nuclear magnetic resonance, The results obtained suggest that the domain assumes a nascent alpha-helical structure in aqueous solution, which is stabilized under hydrophobic environments.
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页码:7639 / 7645
页数:7
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