PMID- 10595557
OWN - NLM
STAT- MEDLINE
DCOM- 20000107
LR  - 20181113
IS  - 0961-8368 (Print)
IS  - 1469-896X (Electronic)
IS  - 0961-8368 (Linking)
VI  - 8
IP  - 11
DP  - 1999 Nov
TI  - A common mechanism for recombinant human NGF, BDNF, NT-3, and murine NGF slow 
      unfolding.
PG  - 2513-8
AB  - The recombinant human nerve growth factor (hNGF), brain-derived neurotrophic 
      factor (BDNF), neurotrophin-3 (NT-3), neurotrophin 4/5 (NT4/5), and murine NGF 
      (mNGF) dimers all undergo rapid unfolding and dissociation to monomer in GdnHCl. 
      Fluorescence spectroscopy, reversed-phase high-performance liquid chromatography, 
      and size-exclusion chromatography were used to show that this monomer M1 converts 
      slowly to a more fully unfolded monomer, M2, by a first order process with 
      half-lives of 22, 2.5, 1.6, and 0.73 h for hNGF, mNGF, NT-3, and BDNF, 
      respectively, at 25 degrees C. Linear Arrhenius plots for the conversion of M1 to 
      M2 yielded activation energies of 27, 22, 24, and 24 kcal/mol for hNGF, mNGF, 
      NT-3, and BDNF, respectively. The refolding of these neurotrophins from 5 M 
      GdnHCl was also first order with NT-3 the slowest to refold and BDNF the fastest. 
      Threading of the N-terminus out through the cystine-knot loop present in each of 
      these proteins is proposed as the slow step in unfolding. The number of amino 
      acids in the cystine-knot loop (14 for hNGF, mNGF, NT-3, and BDNF; 21 for NT4/5), 
      and the number and position of the proline residues in this loop (2 for hNGF; 1 
      for mNGF, NT-3, BDNF, and NT4/5) correlate with the relative rates of unfolding. 
      The smaller the loop and the greater the number of prolines, the more hindered 
      and slower the unfolding.
FAU - De Young, L R
AU  - De Young LR
AD  - Department of Pharmaceutical Research and Development, Genentech, Inc., South San 
      Francisco, California 94080, USA. Ldeyoung@angiogenix.com
FAU - Schmelzer, C H
AU  - Schmelzer CH
FAU - Burton, L E
AU  - Burton LE
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Protein Sci
JT  - Protein science : a publication of the Protein Society
JID - 9211750
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Nerve Growth Factors)
RN  - 0 (Neurotrophin 3)
RN  - 0 (Recombinant Proteins)
RN  - JU58VJ6Y3B (Guanidine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Brain-Derived Neurotrophic Factor/*chemistry/metabolism
MH  - Chromatography, Gel
MH  - Chromatography, High Pressure Liquid
MH  - Guanidine
MH  - Humans
MH  - Kinetics
MH  - Macromolecular Substances
MH  - Mice
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Nerve Growth Factors/*chemistry/metabolism
MH  - Neurotrophin 3/*chemistry/metabolism
MH  - Protein Conformation
MH  - *Protein Denaturation
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Thermodynamics
PMC - PMC2144210
EDAT- 1999/12/14 00:00
MHDA- 1999/12/14 00:01
PMCR- 2000/05/01
CRDT- 1999/12/14 00:00
PHST- 1999/12/14 00:00 [pubmed]
PHST- 1999/12/14 00:01 [medline]
PHST- 1999/12/14 00:00 [entrez]
PHST- 2000/05/01 00:00 [pmc-release]
AID - 10.1110/ps.8.11.2513 [doi]
PST - ppublish
SO  - Protein Sci. 1999 Nov;8(11):2513-8. doi: 10.1110/ps.8.11.2513.