Dissecting NGF interactions with TrkA and p75 receptors by structural and functional studies of an anti-NGF neutralizing antibody.
The anti-nerve growth factor (NGF) monoclonal antibody alphaD11 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 alphaD11 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 alphaD11 Fab fragment was solved at 1.7 A resolution. A computational docking model of the alphaD11 Fab-NGF complex, based on epitope mapping using a pool of 44 NGF mutants and experimentally validated by small-angle X-ray scattering, provided the structural basis for identifying the residues involved in alphaD11 Fab binding. The present study pinpoints loop II of NGF to be an important structural determinant for NGF biological activity mediated by TrkA receptor.
Covaceuszach
Sonia
S
Lay Line Genomics SpA, via di Castel Romano 100, I-00128 Rome, Italy.
Cassetta
Alberto
A
Konarev
Petr V
PV
Gonfloni
Stefania
S
Rudolph
Rainer
R
Svergun
Dmitri I
DI
Lamba
Doriano
D
Cattaneo
Antonino
A
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006801:Humans; D006860:Hydrogen Bonding; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000737:chemistry* / Q000378:metabolism*; D009500:Neutralization Tests; D017433:Protein Structure, Secondary; D051381:Rats; D020917:Receptor, trkA / Q000737:chemistry* / Q000378:metabolism*; D053838:Scattering, Small Angle; D013329:Structure-Activity Relationship; D014961:X-Ray Diffraction
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D020932:Nerve Growth Factor; D020917:Receptor, trkA