Dissecting NGF interactions with TrkA and p75 receptors by structural and functional studies of an anti-NGF neutralizing antibody

被引：36

作者：

Covaceuszach, Sonia ^{[2
,3
]}

Cassetta, Alberto ^{[4
]}

Konarev, Petr V. ^{[5
,6
]}

Gonfloni, Stefania ^{[3
,7
]}

Rudolph, Rainer ^{[8
]}

Svergun, Dmitri I. ^{[5
,6
]}

Lamba, Doriano ^{[4
]}

Cattaneo, Antonino ^{[1
,3
]}

机构：

[1] European Brain Res Inst, I-00143 Rome, Italy

[2] Lay Line Geonom SpA, I-00128 Rome, Italy

[3] Scuola Int Super Studi Avanzati, Neurosci Program, I-34012 Trieste, Italy

[4] CNR, Inst Cristallog, I-34012 Trieste, Italy

[5] European Mol Biol Lab, Hamburg Outstn, D-22603 Hamburg, Germany

[6] Russian Acad Sci, Inst Crystallog, Moscow 117333, Russia

[7] Univ Roma Tor Vergata, Dept Biol, I-00133 Rome, Italy

[8] Univ Halle Wittenberg, Inst Biotechnol, D-06120 Halle, Germany

来源：

JOURNAL OF MOLECULAR BIOLOGY | 2008年 / 381卷 / 04期

关键词：

anti-NGF antibody; NGF receptors; epitope mapping; SAXS; docking;

D O I：

10.1016/j.jmb.2008.06.008

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 anti-nerve growth factor (NGF) monoclonal antibody alpha D11 is a potent antagonist that neutralizes the biological functions of its antigen in vivo. NGF antagonism is expected to be a highly effective and safe therapeutic approach in many pain states. A comprehensive functional and structural analysis of alpha D11 monoclonal antibody was carried out, showing its ability to neutralize NGF binding to either tropomyosine receptor kinase A (TrkA) or p75 receptors. The 3-D structure of the alpha D11 Fab fragment was solved at 1.7 angstrom resolution. A computational docking model of the alpha D11 Fab-NGF complex, based on epitope mapping using a pool of 44 NGF mutants and experimentally validated by small-angle Xray scattering, provided the structural basis for identifying the residues involved in alpha D11 Fab binding. The present study pinpoints loop II of NGF to be an important structural determinant for NGF biological activity mediated by TrkA receptor. (C) 2008 Elsevier Ltd. All rights reserved.

引用

页码：881 / 896

页数：16

共 71 条

[1] Characterization of symmetric complexes of nerve growth factor and the ectodomain of the pan-neurotrophin receptor, p75NTR [J].

Aurikko, JP ;

Ruotolo, BT ;

Grossmann, JG ;

Moncrieffe, MC ;

Stephens, E ;

Leppänen, VM ;

Robinson, CV ;

Saarma, M ;

Bradshaw, RA ;

Blundell, TL .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (39) :33453-33460

[2]

Balestra E, 2001, J BIOL REG HOMEOS AG, V15, P272

[3] Structure of the Fab fragment of a monoclonal antibody specific for carcinoembryonic antigen [J].

Banfield, MJ ;

King, DJ ;

Mountain, A ;

Brady, RL .

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1996, 52 :1107-1113

[4] MONOCLONAL-ANTIBODIES TO NERVE GROWTH-FACTOR AFFECT THE POSTNATAL-DEVELOPMENT OF THE VISUAL-SYSTEM [J].

BERARDI, N ;

CELLERINO, A ;

DOMENICI, L ;

FAGIOLINI, M ;

PIZZORUSSO, T ;

CATTANEO, A ;

MAFFEI, L .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (02) :684-688

[5] The Protein Data Bank [J].

Berman, HM ;

Westbrook, J ;

Feng, Z ;

Gilliland, G ;

Bhat, TN ;

Weissig, H ;

Shindyalov, IN ;

Bourne, PE .

NUCLEIC ACIDS RESEARCH, 2000, 28 (01) :235-242

[6] Neurotrophins: key regulators of cell fate and cell shape in the vertebrate nervous system [J].

Bibel, M ;

Barde, YA .

GENES & DEVELOPMENT, 2000, 14 (23) :2919-2937

[7]

BOTHWELL M, 1995, ANNU REV NEUROSCI, V18, P223, DOI 10.1146/annurev.ne.18.030195.001255

[8] DATA ACQUISITION-SYSTEMS FOR LINEAR AND AREA X-RAY-DETECTORS USING DELAY-LINE READOUT [J].

BOULIN, CJ ;

KEMPF, R ;

GABRIEL, A ;

KOCH, MHJ .

NUCLEAR INSTRUMENTS & METHODS IN PHYSICS RESEARCH SECTION A-ACCELERATORS SPECTROMETERS DETECTORS AND ASSOCIATED EQUIPMENT, 1988, 269 (01) :312-320

[9]

BRUGNER AT, 1998, ACTA CRYSTALLOGR D, V54, P905

[10] Structural changes in a cryo-cooled protein crystal owing to radiation damage [J].

Burmeister, WP .

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2000, 56 :328-341

← 1 2 3 4 5 6 7 8 →