PMID- 938638
OWN - NLM
STAT- MEDLINE
DCOM- 19760925
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 15
IP  - 12
DP  - 1976 Jun 15
TI  - Folding and interaction of subunits at the antibody combining site.
PG  - 2706-10
AB  - The Fv fragment derived from mouse myeloma protein 315 possessing 
      anti-dinitrophenyl (DNP) activity, is composed of two subunits, the peptide chain 
      VL and VH. In 8 M urea there is a complete dissociation of VL and VH and an 
      approximately twofold increase in the fluorescence emission of Fv with a 
      characteristic red shift of 11 nm. Upon dilution of Fv from 8 M urea into neutral 
      buffer full regain of activity was observed, concomitant with regain of native 
      fluorescence spectrum. The decrease in fluorescence upon dilution from 8 M urea 
      was used to follow the renaturation process of Fv. At relatively high protein 
      concentration (2.5 x 10(-6) M) two steps were observed during renaturation: a 
      fast one, which is completed in less than 30 s, and a slower step, which proceeds 
      for approximately 20 min. The fast process represents the refolding and 
      association of VL and VH to form an active FV, whereas the slow step is 
      attributed to the formation of "incorrect" associates between VL and VH which 
      slowly reshuffle to the thermodynamically stable active FV. Indeed, at low 
      protein concentration (1.5 x 10(-8 M) only the fast step is observed and 
      renaturation is completed in less than 30s. The presence of hapten does not 
      affect the rate of renaturation of FV. Reoxidation of FV completely reduced in 8 
      M urea was also found to yield a fully active Fv. Since either VL or VH have only 
      one intrachain disulfide bond, reoxidation was performed at high protein 
      concentration (3 mg/ml) in 8 M urea followed by dilution into neutral buffer. 
      This demonstrates that variable domains not only exist in immunoglobulin 
      structure but can also fold correctly independent of the rest of the peptide 
      chains.
FAU - Hochman, J
AU  - Hochman J
FAU - Gavish, M
AU  - Gavish M
FAU - Inbar, D
AU  - Inbar D
FAU - Givol, D
AU  - Givol D
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Dinitrophenols)
RN  - 0 (Immunoglobulin Fab Fragments)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Myeloma Proteins)
RN  - 8W8T17847W (Urea)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - *Binding Sites, Antibody
MH  - Dinitrophenols
MH  - *Immunoglobulin Fab Fragments
MH  - Kinetics
MH  - Macromolecular Substances
MH  - Mice
MH  - *Myeloma Proteins
MH  - Protein Binding
MH  - Protein Conformation
MH  - Protein Denaturation
MH  - Spectrometry, Fluorescence
MH  - Urea
EDAT- 1976/06/15 00:00
MHDA- 1976/06/15 00:01
CRDT- 1976/06/15 00:00
PHST- 1976/06/15 00:00 [pubmed]
PHST- 1976/06/15 00:01 [medline]
PHST- 1976/06/15 00:00 [entrez]
AID - 10.1021/bi00657a034 [doi]
PST - ppublish
SO  - Biochemistry. 1976 Jun 15;15(12):2706-10. doi: 10.1021/bi00657a034.