PMID- 1832555
OWN - NLM
STAT- MEDLINE
DCOM- 19911023
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 30
IP  - 38
DP  - 1991 Sep 24
TI  - Conformations of IgE bound to its receptor Fc epsilon RI and in solution.
PG  - 9125-32
AB  - Previous resonance energy transfer studies suggested that murine immunoglobulin E 
      (IgE) is bent near the junction of its Fc and Fab segments when bound to its 
      high-affinity receptor (Fc epsilon RI) on RBL cells. To examine further the 
      conformations of IgE, both bound to this receptor and in solution, a mutant 
      recombinant IgE (epsilon/C gamma 3*) was prepared that has a cysteine replacing a 
      serine near the C-terminal ends of the heavy chain. The introduced cysteine 
      residues provide a means for specific modification of IgE, and the sulfhydryl 
      groups were selectively labeled with fluorescein-5-maleimide (FM-epsilon/C gamma 
      3*). This IgE also binds a 5-(dimethylamino)naphthalene-1-sulfonyl (DNS) group in 
      the antigen-binding sites. Resonance energy transfer experiments carried out on 
      receptor-bound FM-epsilon/C gamma 3* yielded a distance of 53 A between 
      fluorescein near the C-terminal end of the Fc segment and amphipathic acceptor 
      probes at the membrane surface. The average distance between this C-terminal 
      fluorescein and acceptor eosin-DNS in the antigen-binding sites at the N-terminal 
      ends of the Fab segments was found to be 69 A. These results combine with those 
      from previous structural studies to provide an unprecedented detailed description 
      of the bent geometry of IgE bound to its receptor on the membrane. Energy 
      transfer measured for FM-epsilon/C gamma 3* in solution between fluorescein near 
      the C-terminal end of the Fc segment and eosin-DNS at the N-terminal ends of the 
      Fab segments indicates that the average distance between these probes is about 71 
      A.(ABSTRACT TRUNCATED AT 250 WORDS)
FAU - Zheng, Y
AU  - Zheng Y
AD  - Department of Chemistry, Cornell University, Ithaca, New York 14853.
FAU - Shopes, B
AU  - Shopes B
FAU - Holowka, D
AU  - Holowka D
FAU - Baird, B
AU  - Baird B
LA  - eng
GR  - AI18306/AI/NIAID NIH HHS/United States
GR  - AI22449/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 (Antigens, Differentiation, B-Lymphocyte)
RN  - 0 (Receptors, Fc)
RN  - 0 (Receptors, IgE)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Animals
MH  - Antigens, Differentiation, B-Lymphocyte/chemistry/metabolism
MH  - Binding Sites
MH  - Binding Sites, Antibody
MH  - Cell Membrane/metabolism
MH  - Energy Transfer
MH  - Immunoglobulin E/chemistry/*ultrastructure
MH  - In Vitro Techniques
MH  - Mice
MH  - Protein Conformation
MH  - Receptors, Fc/chemistry/metabolism/*ultrastructure
MH  - Receptors, IgE
MH  - Solubility
EDAT- 1991/09/24 00:00
MHDA- 1991/09/24 00:01
CRDT- 1991/09/24 00:00
PHST- 1991/09/24 00:00 [pubmed]
PHST- 1991/09/24 00:01 [medline]
PHST- 1991/09/24 00:00 [entrez]
AID - 10.1021/bi00102a002 [doi]
PST - ppublish
SO  - Biochemistry. 1991 Sep 24;30(38):9125-32. doi: 10.1021/bi00102a002.