Induction of raft-like domains by a myristoylated NAP-22 peptide and its Tyr mutant

被引:17
作者
Epand, RF
Sayer, BG
Epand, RM [1 ]
机构
[1] McMaster Univ, Dept Biochem & Biomed Sci, Hamilton, ON L8N 3Z5, Canada
[2] McMaster Univ, Dept Chem, Hamilton, ON, Canada
关键词
cholesterol; domains; differential scanning calorimetry; MAS/NMR; phosphatidylinositol (4,5) diphosphate;
D O I
10.1111/j.1742-4658.2005.04612.x
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
The N-terminally myristoylated, 19-amino acid peptide, corresponding to the amino terminus of the neuronal protein NAP-22 (NAP-22 peptide) is a naturally occurring peptide that had been shown by fluorescence to cause the sequestering of a Bodipy-labeled PtdIns(4,5)P2 in a cholesterol-dependent manner. The present work, using differential scanning calorimetry (DSC), extends the observation that formation of a PtdIns(4,5)P2-rich domain is cholesterol dependent and shows that it also leads to the formation of a cholesterol-depleted domain. The PtdIns(4,5)P2 used in the present work is extracted from natural sources and does not contain any label and has the native acyl chain composition. Peptide-induced formation of a cholesterol-depleted domain is abolished when the sole aromatic amino acid, Tyr11 is replaced with a Leu. Despite this, the modified peptide can still sequester PtdIns(4,5)P2 into domains, probably because of the presence of a cluster of cationic residues in the peptide. Cholesterol and PtdIns(4,5)P2 also modulate the insertion of the peptide into the bilayer as revealed by H-1 NOESY MAS/NMR. The intensity of cross peaks between the aromatic protons of the Tyr residue and the protons of the lipid indicate that in the presence of cholesterol there is a change in the nature of the interaction of the peptide with the membrane. These results have important implications for the mechanism by which NAP-22 affects actin reorganization in neurons.
引用
收藏
页码:1792 / 1803
页数:12
相关论文
共 48 条
[1]  
Ames B. N., 1966, METHOD ENZYMOL, V8, P115, DOI DOI 10.1016/0076-6879(66)08014-5
[2]   1H-NMR parameters of common amino acid residues measured in aqueous solutions of the linear tetrapeptides Gly-Gly-X-Ala at pressures between 0.1 and 200 MPa [J].
Arnold, MR ;
Kremer, W ;
Lüdemann, HD ;
Kalbitzer, HR .
BIOPHYSICAL CHEMISTRY, 2002, 96 (2-3) :129-140
[3]   Actin cytoskeleton regulation through modulation of PI(4,5)P2 rafts [J].
Caroni, P .
EMBO JOURNAL, 2001, 20 (16) :4332-4336
[4]   The S4 genome segment of baboon reovirus is bicistronic and encodes a novel fusion-associated small transmembrane protein [J].
Dawe, S ;
Duncan, R .
JOURNAL OF VIROLOGY, 2002, 76 (05) :2131-2140
[5]   Membrane position of a basic aromatic peptide that sequesters phosphatidylinositol 4,5 bisphosphate determined by site-directed spin labeling and high-resolution NMR [J].
Ellena, JF ;
Moulthrop, J ;
Wu, J ;
Rauch, M ;
Jaysinghne, S ;
Castle, JD ;
Cafiso, DS .
BIOPHYSICAL JOURNAL, 2004, 87 (05) :3221-3233
[6]   Specificity of membrane binding of the neuronal protein NAP-22 [J].
Epand, RF ;
Maekawa, S ;
Epand, RM .
JOURNAL OF MEMBRANE BIOLOGY, 2003, 193 (03) :171-176
[7]   Protein-induced formation of cholesterol-rich domains [J].
Epand, RM ;
Maekawa, S ;
Yip, CM ;
Epand, RF .
BIOCHEMISTRY, 2001, 40 (35) :10514-10521
[8]   Cholesterol-dependent partitioning of Ptdlns(4,5)P2 into membrane domains by the N-terminal fragment of NAP-22 (neuronal axonal myristoylated membrane protein of 22 kDa) [J].
Epand, RM ;
Vuong, P ;
Yip, CM ;
Maekawa, S ;
Epand, RF .
BIOCHEMICAL JOURNAL, 2004, 379 :527-532
[9]   Peptide-induced formation of cholesterol-rich domains [J].
Epand, RM ;
Sayer, BG ;
Epand, RF .
BIOCHEMISTRY, 2003, 42 (49) :14677-14689
[10]   Novel properties of cholesterol-dioleoylphosphatidylcholine mixtures [J].
Epand, RM ;
Hughes, DW ;
Sayer, BG ;
Borochov, N ;
Bach, D ;
Wachtel, E .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2003, 1616 (02) :196-208