PMID- 2935182
OWN - NLM
STAT- MEDLINE
DCOM- 19860317
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 24
IP  - 22
DP  - 1985 Oct 22
TI  - Structural mapping of membrane-bound immunoglobulin E-receptor complexes: use of 
      monoclonal anti-IgE antibodies to probe the conformation of receptor-bound IgE.
PG  - 6260-7
AB  - Previous resonance energy-transfer measurements have suggested that 
      immunoglobulin E (IgE) may bend near the junction of its Fc and Fab segments in 
      order to bind to its high-affinity receptor on rat basophilic leukemia cells. In 
      order to test this possibility, two monoclonal antibodies were employed that bind 
      specifically to rat IgE (IgER) when IgER is in solution and when it is bound to 
      receptors on the plasma membrane. The F(ab')2 fragment of one monoclonal (B5) 
      that is specific for the Fab region of IgER was labeled with donor probes and 
      bound to IgER, and the quenching of the fluorescence of these donors due to 
      simultaneous binding of the Fab' fragment of an anti-Fc monoclonal (A2) that was 
      labeled with an acceptor probe at its interchain disulfide bond was measured. 
      Significantly less energy transfer between these probes was observed when IgER 
      was bound to its receptor on membrane vesicles than when it was free in solution, 
      and this result is interpreted in light of other energy-transfer measurements 
      using A2 and B5 that were preferentially labeled near their combining sites with 
      donors and acceptors, respectively, as well as measurements of the distance of 
      closest approach between these sites and the membrane surface. These results 
      along with previous energy-transfer measurements and other biochemical 
      information form the basis for a working model of the conformation and 
      orientation of receptor-bound IgE. This study demonstrates the use of 
      fluorescently labeled monoclonal antibodies as highly selective energy-transfer 
      probes in assessing structures of macromolecular complexes on the plasma 
      membrane.
FAU - Holowka, D
AU  - Holowka D
FAU - Conrad, D H
AU  - Conrad DH
FAU - Baird, B
AU  - Baird B
LA  - eng
GR  - AI18306/AI/NIAID NIH HHS/United States
GR  - AI18610/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
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 (Fluoresceins)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Receptors, Fc)
RN  - 0 (Receptors, IgE)
RN  - 0 (Receptors, Immunologic)
RN  - 0 (Thiocyanates)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - I223NX31W9 (Fluorescein-5-isothiocyanate)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal
MH  - Basophils/immunology
MH  - Cell Membrane/immunology
MH  - Energy Transfer
MH  - Fluorescein-5-isothiocyanate
MH  - Fluoresceins
MH  - Fluorescent Dyes
MH  - Immunoglobulin E/*metabolism
MH  - Kinetics
MH  - Leukemia, Experimental/immunology
MH  - Models, Molecular
MH  - Protein Conformation
MH  - Rats
MH  - Receptors, Fc/isolation & purification/*metabolism
MH  - Receptors, IgE
MH  - Receptors, Immunologic/isolation & purification/*metabolism
MH  - Thiocyanates
EDAT- 1985/10/22 00:00
MHDA- 1985/10/22 00:01
CRDT- 1985/10/22 00:00
PHST- 1985/10/22 00:00 [pubmed]
PHST- 1985/10/22 00:01 [medline]
PHST- 1985/10/22 00:00 [entrez]
AID - 10.1021/bi00343a033 [doi]
PST - ppublish
SO  - Biochemistry. 1985 Oct 22;24(22):6260-7. doi: 10.1021/bi00343a033.