Identification of the high affinity binding site in transforming growth factor-β involved in complex formation with α2-macroglobulin -: Implications regarding the molecular mechanisms of complex formation between α2-macroglobulin and growth factors, cytokines, and hormones

被引:24
作者
Liu, QJ
Ling, TY
Shieh, HS
Johnson, FE
Huang, JS
Huang, SS
机构
[1] St Louis Univ, Sch Med, Dept Biochem & Mol Biol, St Louis, MO 63104 USA
[2] St Louis Univ, Sch Med, Dept Surg, St Louis, MO 63104 USA
[3] Acad Sinica, Inst Biomed Sci, Taipei 11529, Taiwan
[4] Pharmacia Monsanto, Searle Discovery Res, St Louis, MO 63198 USA
关键词
D O I
10.1074/jbc.M105177200
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 biological activities of transforming growth factor-beta isoforms (TGF-beta (1,2)) are known to be modulated by alpha (2)-macroglobulin (alpha M-2). alpha M-2 forms complexes with numerous growth factors, cytokines, and hormones, including TGF-beta. Identification of the binding sites in TGF-beta isoforms responsible for high affinity interaction with a2M many unravel the molecular basis of the complex formation. Here we demonstrate that among nine synthetic pentacosapeptides with overlapping amino acid sequences spanning the entire TGF-beta (1), molecule, the peptide (residues 41-65) containing Trp-52 exhibited the most potent activity in inhibiting the formation of complexes between I-125-TGF-beta (1) and activated alpha M-2 (alpha M-2*) as determined by nondenaturing polyacrylamide gel electrophoresis and by plasma clearance in mice. TGF-beta (2) peptide containing the homologous sequence and Trp-52 was as active as the TGF-beta (1) peptide, whereas the corresponding TGF-beta (3) peptide lacking Trp-52, was inactive. The replacement of the Trp-52 with alanine abolished the inhibitory activities of these peptides. I-125-TGF-beta (3), which lacks Trp-52, bound to a2M* with an affinity lower than that of I-125-TGF-beta (1). Furthermore, unlabeled TGF-beta (3) and the mutant TGF-beta (1)W52A, in which Trp-52 was replaced with alanine, were less potent than unlabeled TGF-P, in blocking I-125-TGF-beta (1) binding to alpha M-2*.TGF-beta (1) and TGF-beta (2) peptides containing Trp-52 were also effective in inhibiting I-125-nerve growth factor binding to a2M*. These results suggest that Trp-52 is involved in high affinity binding of TGF-beta to alpha M-2*. They also imply that TGF-beta and other growth factors/cytokines/hormones may form complexes with a2M* via a common mechanism involving the interactions between topologically exposed Trp and/or other hydrophobic residues and a hydrophobic region in alpha M-2*.
引用
收藏
页码:46212 / 46218
页数:7
相关论文
共 46 条
[1]   TRANSFORMING GROWTH FACTOR-BETA-1 - SECONDARY STRUCTURE AS DETERMINED BY HETERONUCLEAR MAGNETIC-RESONANCE SPECTROSCOPY [J].
ARCHER, SJ ;
BAX, A ;
ROBERTS, AB ;
SPORN, MB ;
OGAWA, Y ;
PIEZ, KA ;
WEATHERBEE, JA ;
TSANG, MLS ;
LUCAS, R ;
ZHENG, BL ;
WENKER, J ;
TORCHIA, DA .
BIOCHEMISTRY, 1993, 32 (04) :1164-1171
[2]   TRANSFORMING GROWTH FACTOR-BETA-1 - NMR SIGNAL ASSIGNMENTS OF THE RECOMBINANT PROTEIN EXPRESSED AND ISOTOPICALLY ENRICHED USING CHINESE-HAMSTER OVARY CELLS [J].
ARCHER, SJ ;
BAX, A ;
ROBERTS, AB ;
SPORN, MB ;
OGAWA, Y ;
PIEZ, KA ;
WEATHERBEE, JA ;
TSANG, MLS ;
LUCAS, R ;
ZHENG, BL ;
WENKER, J ;
TORCHIA, DA .
BIOCHEMISTRY, 1993, 32 (04) :1152-1163
[3]   INTERACTION OF ALPHA2-MACROGLOBULIN WITH PROTEINASES - CHARACTERISTICS AND SPECIFICITY OF REACTION, AND A HYPOTHESIS CONCERNING ITS MOLECULAR MECHANISM [J].
BARRETT, AJ ;
STARKEY, PM .
BIOCHEMICAL JOURNAL, 1973, 133 (04) :709-&
[4]  
BIRKENMEIER G, 1994, METHOD ENZYMOL, V228, P264
[5]   ALPHA(2)-MACROGLOBULIN - A MULTIFUNCTIONAL BINDING AND TARGETING PROTEIN WITH POSSIBLE ROLES IN IMMUNITY AND AUTOIMMUNITY [J].
BORTH, W .
BIOLOGY OF ALPHA2-MACROGLOBULIN, ITS RECEPTOR, AND RELATED PROTEINS, 1994, 737 :267-272
[6]   ALPHA(2)-MACROGLOBULIN, A MULTIFUNCTIONAL BINDING-PROTEIN WITH TARGETING CHARACTERISTICS [J].
BORTH, W .
FASEB JOURNAL, 1992, 6 (15) :3345-3353
[7]   CHARACTERIZATION OF DISTINCT FUNCTIONAL DOMAINS OF TRANSFORMING GROWTH-FACTOR-BETA [J].
BURMESTER, JK ;
QIAN, SW ;
ROBERTS, AB ;
HUANG, A ;
AMATAYAKULCHANTLER, S ;
SUARDET, L ;
ODARTCHENKO, N ;
MADRI, JA ;
SPORN, MB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (18) :8628-8632
[8]   MECHANISM OF INSULIN INCORPORATION INTO ALPHA-2-MACROGLOBULIN - IMPLICATIONS FOR THE STUDY OF PEPTIDE AND GROWTH-FACTOR BINDING [J].
CHU, CT ;
RUBENSTEIN, DS ;
ENGHILD, JJ ;
PIZZO, SV .
BIOCHEMISTRY, 1991, 30 (06) :1551-1560
[9]  
Coté N, 1998, CAN J VET RES, V62, P279
[10]   BINDING OF PLATELET-DERIVED GROWTH FACTOR-BB AND TRANSFORMING GROWTH FACTOR-BETA(1) TO ALPHA(2)-MACROGLOBULIN IN-VITRO AND IN-VIVO - COMPARISON OF RECEPTOR-RECOGNIZED AND NON-RECOGNIZED ALPHA(2)-MACROGLOBULIN CONFORMATIONS [J].
CROOKSTON, KP ;
WEBB, DJ ;
LAMARRE, J ;
GONIAS, SL .
BIOCHEMICAL JOURNAL, 1993, 293 :443-450