PMID- 8399230
OWN - NLM
STAT- MEDLINE
DCOM- 19931124
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 32
IP  - 40
DP  - 1993 Oct 12
TI  - Refolding of brain-derived neurotrophic factor from guanidine hydrochloride: 
      kinetic trapping in a collapsed form which is incompetent for dimerization.
PG  - 10812-8
AB  - We have studied the pathway and kinetics of refolding of recombinant human 
      brain-derived neurotrophic factor (BDNF), which is a very tightly-associated 
      dimer in its native state. When BDNF unfolded in 6 M guanidine hydrochloride is 
      diluted 20-fold into phosphate-buffered saline, a partially folded intermediate 
      is rapidly formed (< 1 min). Circular dichroism and fluorescence spectroscopy 
      show that this intermediate has extensive secondary structure, but no 
      well-defined tertiary structure. Size-exclusion chromatography with light 
      scattering detection shows that it is compact and monomeric, and therefore 
      corresponds to what is often called a "collapsed form" or "molten globule". This 
      collapsed form disappears with a half-time of approximately 30 min, 
      simultaneously with the appearance of native dimers, without accumulation of 
      monomeric species with a native tertiary structure. Remarkably, the monomer-dimer 
      association constant of the collapsed form is approximately 10(10) weaker than 
      the native structure, and it has a low tendency to form large aggregates. Given 
      the very large hydrophobic surface present at the dimer interface of nerve growth 
      factor (and presumably in BDNF), these results indicate that these hydrophobic 
      groups are not exposed in the collapsed form, and that it is therefore quite 
      dissimilar from the native structure. A significant conformational change in the 
      collapsed form is necessary to re-expose these hydrophobic groups to form the 
      dimer interface, making this the rate-limiting step in reaching the native 
      conformation.
FAU - Philo, J S
AU  - Philo JS
AD  - Protein Chemistry Department, Amgen Inc., Amgen Center, Thousand Oaks, California 
      91320.
FAU - Rosenfeld, R
AU  - Rosenfeld R
FAU - Arakawa, T
AU  - Arakawa T
FAU - Wen, J
AU  - Wen J
FAU - Narhi, L O
AU  - Narhi LO
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Guanidines)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Nerve Growth Factors)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Recombinant Proteins)
RN  - JU58VJ6Y3B (Guanidine)
SB  - IM
MH  - Brain-Derived Neurotrophic Factor
MH  - Chromatography, Gel
MH  - Circular Dichroism
MH  - Cloning, Molecular
MH  - Escherichia coli
MH  - Guanidine
MH  - Guanidines/pharmacology
MH  - Humans
MH  - Kinetics
MH  - Macromolecular Substances
MH  - Nerve Growth Factors/*chemistry
MH  - Nerve Tissue Proteins/*chemistry/isolation & purification/metabolism
MH  - *Protein Conformation
MH  - *Protein Folding
MH  - Protein Structure, Secondary
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/chemistry/isolation & purification/metabolism
EDAT- 1993/10/12 00:00
MHDA- 1993/10/12 00:01
CRDT- 1993/10/12 00:00
PHST- 1993/10/12 00:00 [pubmed]
PHST- 1993/10/12 00:01 [medline]
PHST- 1993/10/12 00:00 [entrez]
AID - 10.1021/bi00091a036 [doi]
PST - ppublish
SO  - Biochemistry. 1993 Oct 12;32(40):10812-8. doi: 10.1021/bi00091a036.