PMID- 10684654
OWN - NLM
STAT- MEDLINE
DCOM- 20000323
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 39
IP  - 8
DP  - 2000 Feb 29
TI  - Rebinding of IgE Fabs at haptenated planar membranes: measurement by total 
      internal reflection with fluorescence photobleaching recovery.
PG  - 2042-51
AB  - In previous work, a general analytical theory for ligand rebinding at cell 
      surfaces was developed for a reversible bimolecular reaction between ligands in 
      solution and receptors on a membrane surface [Lagerholm, B. C., and Thompson, N. 
      L. (1998) Biophys. J. 74, 1215-1228]. This theory can be used to predict 
      theoretical forms for data obtained by using total internal reflection with 
      fluorescence photobleaching recovery (TIR-FPR) [Thompson, N. L., Burghardt, T. 
      P., and Axelrod, D. (1981) Biophys. J. 33, 435-454]. Thus, one method by which 
      the rebinding theory can be tested is to use TIR-FPR. In the work described 
      herein, the reversible kinetics of mouse monoclonal anti-dinitrophenyl (DNP) IgE 
      Fabs at substrate-supported planar membranes composed of 25 mol % DNP-conjugated 
      phosphatidylethanolamine and 75 mol % dipalmitoylphosphatidylcholine have been 
      examined by using TIR-FPR. Data were obtained as a function of the Fab solution 
      concentration. Higher Fab concentrations reduce rebinding (and increase the 
      fluorescence recovery rate) because different Fab molecules compete for the same 
      surface-binding sites. Data were also obtained for solutions containing different 
      volume fractions of glycerol. In these measurements, higher glycerol 
      concentrations increase rebinding (and decrease the fluorescence recovery rate) 
      because the solution viscosity is increased and the Fab diffusion coefficient in 
      solution is decreased. The TIR-FPR data were quantitatively compared with 
      theoretical predictions which follow from the general theory for rebinding at the 
      membrane surface. The data were consistent with the theoretical predictions and, 
      therefore, provide experimental verification of the previously developed theory.
FAU - Lagerholm, B C
AU  - Lagerholm BC
AD  - Department of Chemistry, Campus Box 3290, University of North Carolina at Chapel 
      Hill, Chapel Hill, North Carolina 27599-3290, USA.
FAU - Starr, T E
AU  - Starr TE
FAU - Volovyk, Z N
AU  - Volovyk ZN
FAU - Thompson, N L
AU  - Thompson NL
LA  - eng
GR  - GM-37145/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Dinitrobenzenes)
RN  - 0 (Immunoglobulin Fab Fragments)
RN  - 0 (Ligands)
RN  - 0 (Membranes, Artificial)
RN  - 0 (Phospholipids)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/metabolism
MH  - Binding Sites, Antibody
MH  - Dinitrobenzenes/chemistry
MH  - Immunoglobulin E/*metabolism
MH  - Immunoglobulin Fab Fragments/*metabolism
MH  - Kinetics
MH  - Ligands
MH  - Membranes, Artificial
MH  - Mice
MH  - Models, Biological
MH  - Phospholipids/metabolism
MH  - Photochemistry
MH  - Surface Properties
MH  - Time Factors
EDAT- 2000/02/24 09:00
MHDA- 2000/03/25 09:00
CRDT- 2000/02/24 09:00
PHST- 2000/02/24 09:00 [pubmed]
PHST- 2000/03/25 09:00 [medline]
PHST- 2000/02/24 09:00 [entrez]
AID - bi9917434 [pii]
AID - 10.1021/bi9917434 [doi]
PST - ppublish
SO  - Biochemistry. 2000 Feb 29;39(8):2042-51. doi: 10.1021/bi9917434.