SASDAU5: ad11 Fab + NGF (aD11 Fab, Fab + NGF)

Basic information

• Entry: Database: SASBDB / ID: SASDAU5

Sample

ad11 Fab + NGF

• aD11 Fab (protein), Fab, Mus musculus
• NGF (protein), NGF, Mus musculus

• Biological species: Mus musculus (house mouse)
• Citation: Journal: J Mol Biol / Year: 2008; Title: Dissecting NGF interactions with TrkA and p75 receptors by structural and functional studies of an anti-NGF neutralizing antibody.; Authors: Sonia Covaceuszach / Alberto Cassetta / Petr V Konarev / Stefania Gonfloni / Rainer Rudolph / Dmitri I Svergun / Doriano Lamba / Antonino Cattaneo / ; Abstract: 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.

Contact author

• Petr Konarev (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

Models

• Model #140: ; Type: dummy / Software: Dammin / Radius of dummy atoms: 3.40 A / Chi-square value: 4.048144; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: ad11 Fab + NGF / Sample MW: 100 kDa / Entity id: 100 / 101
• Buffer: Name: 30 mM Na-phosphate / pH: 7 / Composition: 75 mM NaCl 75.000 mM

Entity #100

Name: Fab / Type: protein / Description: aD11 Fab / Formula weight: 25 / Num. of mol.: 2 / Source: Mus musculus
Sequence:

DIQMTQSPAS LSASLGETVT IECRASEDIY NALAWYQQKP GKSPQLLIYN TDTLHTGVPS RFSGSGSGTQ YSLKINSLQS EDVASYFCQH YFGYPRTFGG GTKLELKRAD AAPTVSIFPP SSEQLASGGA SVVCLLNNFY PKDISVKWKI DGSERQNGVL DSVTDQDSKD STYSMSSTLT LTKAEYESHN SYTCEVTHKT STSPVVKSFN RGEQVQLKES GPGLVQPSQT LSLTCTVSGF SLTNNNVNWV RQATGRGLEW MGGVWAGGAT DYNSALKSRL TITRDTSKSQ VFLKMHSLQS SDTATYYCAR DGGYSSSTLY AMDAWGQGTT VTVSSASTTA PSVYPLAPGS MVTLGCLVKG YFPEPVTVTW NSGSLASGVH TFPAVLQSGL YTLSSSVTVP ASPWASEAVT CNVAHPASST KVDKKIVPR

Entity #101

Name: NGF / Type: protein / Description: NGF / Formula weight: 25 / Num. of mol.: 2 / Source: Mus musculus
Sequence:

GEFSVCDSVS VWVGDKTTAT DIKGKEVTVL AEVNINNSVF RQYFFETKCR ASNPVESGCR GIDSKHWNSY CTTTHTFVKA LTTDEKQAAW RFIRIDTACV CVLSRKAGEF SVCDSVSVWV GDKTTATDIK GKEVTVLAEV NINNSVFRQY FFETKCRASN PVESGCRGID SKHWNSYCTT THTFVKALTT DEKQAAWRFI RIDTACVCVL SRKA

Experimental information

• Beam: Instrument name: DORIS III X33 / City: Hamburg / 国: Germany / Type of source: X-ray synchrotron
• Detector: Name: MAR 345 Image Plate

Scan

Title: Fab_NGF / Measurement date: Dec 20, 2005 / Storage temperature: 15 °C / Cell temperature: 15 °C / Exposure time: 120 sec. / Number of frames: 2 / Unit: 1/nm /

	Min	Max
Q	0.1517	4.8558

Distance distribution function P(R)

Sofotware P(R): GNOM 4.5a / Number of points: 512 /

	Min	Max
Q	0.1618	4.76
P(R) point	4	515
R	0	16

Result

D max: 16 / Type of curve: single_conc /

	Experimental	Porod
MW	130 kDa	-
Volume	-	255 nm3

	P(R)	Guinier
Forward scattering, I0	2365400	2365600
Radius of gyration, Rg	4.3 nm	4.3 nm