PMID- 4084517
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 Fc receptor bound immunoglobulin E: proximity to the 
      membrane surface of the antibody combining site and another site in the Fab 
      segments.
PG  - 6252-9
AB  - Resonance energy-transfer methods have been used to investigate the structure of 
      immunoglobulin E (IgE) bound to its high-affinity receptor on plasma membrane 
      vesicles derived from rat basophilic leukemia cells. The structural mapping of 
      receptor-bound IgE was initiated in an earlier study [Holowka, D., & Baird, B. 
      (1983) Biochemistry 22, 3475], and it is based on measuring the minimal distance 
      from IgE sites that are selectively labeled with donor probes to a plane of 
      amphipathic acceptors at the membrane surface. This paper describes the use of 
      monoclonal IgE specific for 5-(dimethylamino)naphthalene-1-sulfonyl (DNS) to 
      place a donor probe, DNS-L-Lys, in the antibody combining sites. The distance 
      from these sites to the membrane surface was determined to be greater than 100 A 
      with two different amphipathic acceptor probes. Another site in the Fab segments 
      of monoclonal IgE (anti-dinitrophenyl) could be labeled selectively with 
      N-[4-[7-(diethylamino)-4-methylcoumarin-3-yl]phenyl]maleimide (CPM) in the 
      absence of reducing agents [CPM(-)], and the reaction could not be blocked by 
      prereaction with N-ethylmaleimide. The pattern of CPM(-)-labeled proteolytic 
      fragments and the lack of fluorescence quenching by (trinitrophenyl)lysine in the 
      antibody combining sites suggested the CPM(-)-labeled site to be in the C epsilon 
      1 domain of IgE. The distance between this site on receptor-bound IgE and the 
      membrane surface was determined to be 75-87 A with two different amphipathic 
      acceptors.(ABSTRACT TRUNCATED AT 250 WORDS)
FAU - Baird, B
AU  - Baird B
FAU - Holowka, D
AU  - Holowka D
LA  - eng
GR  - AI18306/AI/NIAID NIH HHS/United States
GR  - AI18610/AI/NIAID NIH HHS/United States
PT  - Journal Article
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 (Immunoglobulin Fab Fragments)
RN  - 0 (Receptors, Fc)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal
MH  - Basophils/enzymology
MH  - Cell Membrane/immunology
MH  - Energy Transfer
MH  - Immunoglobulin E/*metabolism
MH  - Immunoglobulin Fab Fragments/immunology
MH  - Kinetics
MH  - Leukemia, Experimental/immunology
MH  - Rats
MH  - Receptors, Fc/*metabolism
MH  - Spectrometry, Fluorescence
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/bi00343a032 [doi]
PST - ppublish
SO  - Biochemistry. 1985 Oct 22;24(22):6252-9. doi: 10.1021/bi00343a032.