Measurement of the affinity of melittin for zwitterionic and anionic membranes using immobilized lipid biosensors

被引:43
作者
Lee, TH [1 ]
Mozsolits, H [1 ]
Aguilar, MI [1 ]
机构
[1] Monash Univ, Dept Biochem & Mol Biol, Clayton, Vic 3800, Australia
来源
JOURNAL OF PEPTIDE RESEARCH | 2001年 / 58卷 / 06期
关键词
peptide-membrane interaction; surface plasmon resonance; membrane affinity; immobilized lipid chromatography; alpha-helix; circular dichroism;
D O I
10.1034/j.1399-3011.2001.10974.x
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The binding of melittin to zwitterionic dimyristyphosphatidylcholine (DMPC) and anionic dimyristylphosphatidylglycerol (DMPG) was analysed using two different immobilized model membrane systems. The first system used surface plasmon resonance (SPIR), which monitors the real-time binding of peptides to an immobilized hybrid bilayer. SPIR experiments reflected a stronger binding of melittin for DMPG than for DMPC, while kinetic analysis suggested the existence of at least two distinct binding steps. The second lipid biosensor system involved an immobilized phospholipid monolayer covalently attached to a microporous silica surface. The binding of melittin to the immobilized monolayer was then monitored using dynamic elution chromatography with varied methanol concentrations to analyse the binding of melittin to DMPC and DMPG, The nonlinear binding behaviour observed for melittin with the phosphatidylcholine (PC) and phosphatidylglycerol (PG) monolayers compared with the linear retention plots and Gaussian peak shapes observed for the control molecule demonstrated that melittin undergoes significant conformational and orientational changes upon binding to the immobilized PC and PG ligands. The dependence of log k' on per cent methanol also demonstrated a bimodal interaction whereby hydrophobic forces predominated at higher temperatures and methanol concentrations, while other forces, presumably electrostatic in nature, also made a contribution to the affinity of the peptides for the lipid monolayer, particularly at lower temperatures. The complementary use of these two lipid biosensors thus allows the role of hydrophobic and electrostatic forces in peptide-membrane interactions to be studied.
引用
收藏
页码:464 / 476
页数:13
相关论文
共 42 条
[1]   MELITTIN BINDING TO MIXED PHOSPHATIDYLGLYCEROL PHOSPHATIDYLCHOLINE MEMBRANES [J].
BESCHIASCHVILI, G ;
SEELIG, J .
BIOCHEMISTRY, 1990, 29 (01) :52-58
[2]   Lipid-induced conformation and lipid-binding properties of cytolytic and antimicrobial peptides: determination and biological specificity [J].
Blondelle, SE ;
Lohner, K ;
Aguilar, MI .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1999, 1462 (1-2) :89-108
[3]   INFLUENCE OF TRYPTOPHAN RESIDUES ON MELITTINS HEMOLYTIC-ACTIVITY [J].
BLONDELLE, SE ;
SIMPKINS, LR ;
PEREZPAYA, E ;
HOUGHTEN, RA .
BIOCHIMICA ET BIOPHYSICA ACTA, 1993, 1202 (02) :331-336
[4]   DETERMINATION OF HELIX AND BETA-FORM OF PROTEINS IN AQUEOUS-SOLUTION BY CIRCULAR-DICHROISM [J].
CHEN, YH ;
YANG, JT ;
CHAU, KH .
BIOCHEMISTRY, 1974, 13 (16) :3350-3359
[5]   Binding of steroidogenic acute regulatory protein to synthetic membranes suggests an active molten globule [J].
Christensen, K ;
Bose, HS ;
Harris, FM ;
Miller, WL ;
Bell, JD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (20) :17044-17051
[6]   Binding of glycopeptide antibiotics to a model of a vancomycin-resistant bacterium [J].
Cooper, MA ;
Williams, DH .
CHEMISTRY & BIOLOGY, 1999, 6 (12) :891-899
[7]   Kinetic analysis of antibody-antigen interactions at a supported lipid monolayer [J].
Cooper, MA ;
Williams, DH .
ANALYTICAL BIOCHEMISTRY, 1999, 276 (01) :36-47
[8]   A vesicle capture sensor chip for kinetic analysis of interactions with membrane-bound receptors [J].
Cooper, MA ;
Hansson, A ;
Löfås, S ;
Williams, DH .
ANALYTICAL BIOCHEMISTRY, 2000, 277 (02) :196-205
[9]  
Gazzara JA, 1997, J LIPID RES, V38, P2134
[10]   Modulation of tryptophan environment in membrane-bound melittin by negatively charged phospholipids: Implications in membrane organization and function [J].
Ghosh, AK ;
Rukmini, R ;
Chattopadhyay, A .
BIOCHEMISTRY, 1997, 36 (47) :14291-14305