A phage display screen and binding studies with acetylated low density lipoprotein provide evidence for the importance of the scavenger receptor cysteine-rich (SRCR) domain in the ligand-binding function of MARCO

被引:26
作者
Chen, YY
Sankala, M
Ojala, JRM
Sun, Y
Tuuttila, A
Isenman, DE
Tryggvason, K
Pikkarainen, T [1 ]
机构
[1] Karolinska Inst, Dept Med Biochem & Biophys, Div Matrix Biol, S-17177 Stockholm, Sweden
[2] Univ Toronto, Dept Biochem, Toronto, ON M55 1A8, Canada
关键词
D O I
10.1074/jbc.M513628200
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
MARCO is a class A scavenger receptor capable of binding both Gram-negative and -positive bacteria. Using the surface plasmon resonance technique, we show here that a recombinant, soluble form of MARCO, sMARCO, binds the major Gram-negative and -positive bacterial surface components, lipopolysaccharide and lipoteichoic acid. Yet, the interaction of these two polyanions with sMARCO is of much lower affinity than that of polyinosinic acid, a polyanionic inhibitor of bacterial binding to MARCO. To further elucidate the ligand-binding functions of MARCO, we performed a phage display screen with sMARCO. The screening resulted in the enrichment of only a handful of phage clones. Contrary to expectations, no polyanionic peptides, but only those with a predominantly hydrophobic nature, were enriched. One peptide, VRWGSFAAWL, was displayed on two-thirds of the phages recovered after four rounds of screening. The VRWGSFAAWL phage-sMARCO interaction had significantly slower dissociation kinetics than that between sMARCO and lipopolysaccharide or lipoteichoic acid. Further work with this phage, and the second most enriched phage, displaying the peptide RLNWAWWLSY, demonstrated that both peptides bind to the SRCR domain of MARCO, and that they probably bind to the same site. Data base searches suggested that the VRWGSFAAWL peptide represents complement component C4, but we could not convincingly confirm this suggestion. A study with chimeric scavenger receptors indicated that even minor sequence changes in the MARCO scavenger receptor cysteine-rich (SRCR) domain can have profound effects on the binding of the prototypic scavenger receptor ligand, acetylated low density lipoprotein. As shown by differential binding of glutathione S-transferase-VRWGSFAAWL, these differences were very likely due to conformational changes. These findings led to experiments that demonstrated a crucial role of the SRCR domain for acetylated low density lipoprotein binding in MARCO. Thus, our results strengthen the notion that the SRCR domain is the major ligand-binding domain in MARCO. Furthermore, they suggest that the domain may contain multiple ligand-binding sites.
引用
收藏
页码:12767 / 12775
页数:9
相关论文
共 37 条
[1]   Stabilizing nonpolar/polar side-chain interactions in the α-helix [J].
Andrew, CD ;
Penel, S ;
Jones, GR ;
Doig, AJ .
PROTEINS-STRUCTURE FUNCTION AND GENETICS, 2001, 45 (04) :449-455
[2]   The scavenger receptor MARCO is required for lung defense against pneumococcal pneumonia and inhaled particles [J].
Arredouani, M ;
Yang, ZP ;
Ning, YY ;
Qin, GZ ;
Soininen, R ;
Tryggvason, K ;
Kobzik, L .
JOURNAL OF EXPERIMENTAL MEDICINE, 2004, 200 (02) :267-272
[3]  
ASHKENAS J, 1993, J LIPID RES, V34, P983
[4]   Critical aggregation concentrations of gram-negative bacterial lipopolysaccharides (LPS) [J].
Aurell, CA ;
Wistrom, AO .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 253 (01) :119-123
[5]   Identification of uteroglobin-related protein 1 and macrophage scavenger receptor with collagenous structure as a lung-specific ligand-receptor pair [J].
Bin, LH ;
Nielson, LD ;
Liu, XQ ;
Mason, RJ ;
Shu, HB .
JOURNAL OF IMMUNOLOGY, 2003, 171 (02) :924-930
[6]   Arginine residues in domain V have a central role for bacteria-binding activity of macrophage scavenger receptor MARCO [J].
Brännström, A ;
Sankala, M ;
Tryggvason, K ;
Pikkarainen, T .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2002, 290 (05) :1462-1469
[7]   αvβ5 integrin-dependent programmed cell death triggered by a peptide mimic of annexin V [J].
Cardó-Vila, M ;
Arap, W ;
Pasqualini, R .
MOLECULAR CELL, 2003, 11 (05) :1151-1162
[8]   THE TYPE-I MACROPHAGE SCAVENGER RECEPTOR BINDS TO GRAM-POSITIVE BACTERIA AND RECOGNIZES LIPOTEICHOIC ACID [J].
DUNNE, DW ;
RESNICK, D ;
GREENBERG, J ;
KRIEGER, M ;
JOINER, KA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (05) :1863-1867
[9]   A vascular endothelial growth factor high affinity receptor 1-specific peptide with antiangiogenic activity identified using a phage display peptide library [J].
El-Mousawi, M ;
Tchistiakova, L ;
Yurchenko, L ;
Pietrzynski, G ;
Moreno, M ;
Stanimirovic, D ;
Ahmad, D ;
Alakhov, V .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (47) :46681-46691
[10]   Structure of the human macrophage MARCO receptor and characterization of its bacteria-binding region [J].
Elomaa, O ;
Sankala, M ;
Pikkarainen, T ;
Bergmann, U ;
Tuuttila, A ;
Raatikainen-Ahokas, A ;
Sariola, H ;
Tryggvason, K .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (08) :4530-4538