Equilibrium denaturation studies of mouse beta-nerve growth factor.


Abstract

Equilibrium denaturation of dimeric mouse beta-nerve growth factor (beta-NGF) has been studied by monitoring changes in the protein's spectroscopic characteristics. Denaturation of beta-NGF in guanidine hydrochloride and urea resulted in an altered intrinsic fluorescence emission spectrum, fluorescence depolarization, and diminished negative circular dichroism. Native-like spectroscopic properties and specific biological activity are restored when denaturant is diluted from unfolded samples, demonstrating that this process is fully reversible. However, refolding of denatured beta-NGF is dependent on the three disulfide bonds present in the native protein and does not readily occur when the disulfide bonds are reduced. Graphical analysis and nonlinear least-squares fitting of beta-NGF denaturation data demonstrate that denaturation is dependent on the concentration of beta-NGF and is consistent with a two-state model involving native dimer and denatured monomer (N2 = 2D). The conformational stability of mouse beta-NGF calculated according to this model is 19.3 +/- 1.1 kcal/mol in 100 mM sodium phosphate at pH 7. Increasing the hydrogen ion concentration resulted in a 25% decrease in beta-NGF stability at pH 4 relative to pH 7. Study holds ProTherm entries: 10369, 10370, 10371, 10372, 10373, 10374 Extra Details: beta-nerve growth factor; denaturation; fluorescence; protein structure

Submission Details

ID: SyGQaG7Q4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:40 p.m.

Version: 1

Publication Details
Timm DE;Neet KE,Protein Sci. (1992) Equilibrium denaturation studies of mouse beta-nerve growth factor. PMID:1304906
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
5LSD 2017-07-05 recombinant mouse Nerve Growth Factor
1WWW 1999-09-15 2.2 NGF IN COMPLEX WITH DOMAIN 5 OF THE TRKA RECEPTOR
1BET 1994-05-31 2.3 NEW PROTEIN FOLD REVEALED BY A 2.3 ANGSTROM RESOLUTION CRYSTAL STRUCTURE OF NERVE GROWTH FACTOR
4EAX 2012-09-12 2.3 Mouse NGF in complex with Lyso-PS
1SG1 2004-06-01 2.4 Crystal Structure of the Receptor-Ligand Complex between Nerve Growth Factor and the Common Neurotrophin Receptor p75
4ZBN 2015-06-10 2.45 Non-helical DNA Triplex Forms a Unique Aptamer Scaffold for High Affinity Recognition of Nerve Growth Factor
4EDW 2014-04-02 2.48 Nerve Growth Factor in Complex with Fab from humanized version of mouse mAb 911 (tanezumab)
4EDX 2014-04-02 2.5 Nerve Growth Factor in Complex with Fab from mouse mAb 911
1BTG 1996-03-08 2.5 CRYSTAL STRUCTURE OF BETA NERVE GROWTH FACTOR AT 2.5 A RESOLUTION IN C2 SPACE GROUP WITH ZN IONS BOUND
4XPJ 2015-07-15 2.61 Crystal structure of Nerve growth factor in complex with lysophosphatidylinositol
1SGF 1998-05-27 3.15 CRYSTAL STRUCTURE OF 7S NGF: A COMPLEX OF NERVE GROWTH FACTOR WITH FOUR BINDING PROTEINS (SERINE PROTEINASES)
5JZ7 2017-01-11 3.4 NGF IN COMPLEX WITH MEDI578 scFv
2IFG 2007-02-13 3.4 Structure of the extracellular segment of human TRKA in complex with nerve growth factor
3IJ2 2010-01-12 3.75 Ligand-receptor structure
6FFY 2018-08-01 3.9 Structure of the mouse SorCS2-NGF complex

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
93.8 Beta-nerve growth factor P20675 NGF_MASNA
96.3 Beta-nerve growth factor P25427 NGF_RAT
100.0 Beta-nerve growth factor P01139 NGF_MOUSE
90.6 Beta-nerve growth factor P13600 NGF_BOVIN
92.5 Beta-nerve growth factor Q5ISB0 NGF_SAIBB
92.5 Beta-nerve growth factor Q9N2E9 NGF_PONPY
92.5 Beta-nerve growth factor Q29074 NGF_PIG
92.5 Beta-nerve growth factor Q9N2F1 NGF_PANTR
92.5 Beta-nerve growth factor P01138 NGF_HUMAN
92.5 Beta-nerve growth factor Q9N2F0 NGF_GORGO