Action of human group IIa secreted phospholipase A2 on cell membranes -: Vesicle but not heparinoid binding determines rate of fatty acid release by exogenously added enzyme

被引:85
作者
Koduri, RS
Baker, SF
Snitko, Y
Han, SK
Cho, WH
Wilton, DC
Gelb, MH
机构
[1] Univ Washington, Dept Chem, Seattle, WA 98195 USA
[2] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
[3] Univ Southampton, Dept Biochem, Southampton SO16 7PX, Hants, England
[4] Univ Illinois, Dept Chem, Chicago, IL 60607 USA
关键词
D O I
10.1074/jbc.273.48.32142
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
Human group IIa phospholipase A(2) (hIIa-PLA2) is a highly basic protein that is secreted hom a number of cells during inflammation and may play a role in arachidonate liberation and in destruction of invading bacteria. It has been proposed that rodent group IIa PLA, is anchored to cell surfaces via attachment to heparan sulfate proteoglycan and that this interaction facilitates lipolysis. hIIa-PLA2 contains 13 lysines, 2 histidines, and 10 arginines that fall into 10 clusters. A panel of 26 hIIa-PLA2 mutants were prepared in which 1-4 basic residues in each cluster were changed to glutamate or aspartate (charge reversal). A detailed analysis of the affinities of these mutants for anionic vesicles and for heparin and heparan sulfate in vitro and of the specific activities of these proteins for hydrolysis of vesicles in vitro and of living cell. membranes reveal the following trends: 1) the affinity of hIIa-PLA2 for heparin and heparan sulfate is modulated not by a highly localized site of basic residues but by diffuse sites that partially overlap with the interfacial binding site. In contrast, only those residues on the interfacial binding site of hIIa-PLA2 are involved in binding to membranes; 2) the relative ability of these mutants to hydrolyze cellular phospholipids when enzymes were added exogenously to CHO-T(1, NIH-3T3, and RAW 264.7 cells correlates with their relative in vitro affinity for vesicles and not with their affinity for heparin and heparan sulfate. 3) The rates of exogenous hIIa-PLA2-cataIyzed fatty acid release from wild type CHO-K1 cells and two mutant lines, one lacking glycosaminoglycan and one lacking heparan sulfate, were similar. Thus basic residues that modulate interfacial binding are important for plasma membrane fatty acid release by exogenously added hIIa-PLA2. Binding of hIIa-PLA2 to cell surface heparan sulfate does not modulate plasma membrane phospholipid hydrolysis by exogenously added hIIa-PLA2.
引用
收藏
页码:32142 / 32153
页数:12
相关论文
共 61 条
  • [31] STRUCTURE OF CDNA CODING FOR RAT PLATELET PHOSPHOLIPASE-A2
    KOMADA, M
    KUDO, I
    MIZUSHIMA, H
    KITAMURA, N
    INOUE, K
    [J]. JOURNAL OF BIOCHEMISTRY, 1989, 106 (04) : 545 - 547
  • [32] KRAMER RM, 1989, J BIOL CHEM, V264, P5768
  • [33] A SINGLE MUTATION AFFECTS BOTH N-ACETYLGLUCOSAMINYLTRANSFERASE AND GLUCURONOSYLTRANSFERASE ACTIVITIES IN A CHINESE-HAMSTER OVARY CELL MUTANT DEFECTIVE IN HEPARAN-SULFATE BIOSYNTHESIS
    LIDHOLT, K
    WEINKE, JL
    KISER, CS
    LUGEMWA, FN
    BAME, KJ
    CHEIFETZ, S
    MASSAGUE, J
    LINDAHL, U
    ESKO, JD
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (06) : 2267 - 2271
  • [34] Docking phospholipase A2 on membranes using electrostatic potential-modulated spin relaxation magnetic resonance
    Lin, Y
    Nielsen, R
    Murray, D
    Hubbell, WL
    Mailer, C
    Robinson, BH
    Gelb, MH
    [J]. SCIENCE, 1998, 279 (5358) : 1925 - 1929
  • [35] MA YH, 1994, J LIPID RES, V35, P2049
  • [36] THE SECRETORY PHOSPHOLIPASE-A2 GENE IS A CANDIDATE FOR THE MOM1 LOCUS, A MAJOR MODIFIER OF APC(MIN)-INDUCED INTESTINAL NEOPLASIA
    MACPHEE, M
    CHEPENIK, KP
    LIDDELL, RA
    NELSON, KK
    SIRACUSA, LD
    BUCHBERG, AM
    [J]. CELL, 1995, 81 (06) : 957 - 966
  • [37] MARSHALL LA, 1995, J PHARMACOL EXP THER, V274, P1254
  • [38] PURIFICATION OF RABBIT PLATELET SECRETORY PHOSPHOLIPASE-A2 AND ITS CHARACTERISTICS
    MIZUSHIMA, H
    KUDO, I
    HORIGOME, K
    MURAKAMI, M
    HAYAKAWA, M
    KIM, DK
    KONDO, E
    TOMITA, M
    INOUE, K
    [J]. JOURNAL OF BIOCHEMISTRY, 1989, 105 (04) : 520 - 525
  • [39] Type II secretory phospholipase A(2) associated with cell surfaces via C-terminal heparin-binding lysine residues augments stimulus-initiated delayed prostaglandin generation
    Murakami, M
    Nakatani, Y
    Kudo, I
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (47) : 30041 - 30051
  • [40] MURAKAMI M, 1993, J IMMUNOL, V151, P5675