PMID- 9398286
OWN - NLM
STAT- MEDLINE
DCOM- 19980127
LR  - 20061115
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 36
IP  - 50
DP  - 1997 Dec 16
TI  - Identification of contact residues in the IgE binding site of human 
      FcepsilonRIalpha.
PG  - 15579-88
AB  - The high-affinity receptor for immunoglobulin E (IgE), FcepsilonRI, is an 
      alphabetagamma2 tetramer found on mast cells, basophils, and several other types 
      of immune effector cells. The interaction of IgE with the alpha-subunit of 
      FcepsilonRI is central to the pathogenesis of allergy. Detailed knowledge of the 
      mode of interaction of FcepsilonRI with IgE may facilitate the development of 
      inhibitors for general use in the treatment of allergic disease. To this end we 
      have performed site-directed mutagenesis on a soluble form of the FcepsilonRI 
      alpha-chain (sFcepsilonRIalpha). The effects of four mutations in the second 
      immunoglobulin-like domain of sFcepsilonRIalpha upon the kinetics of binding to 
      IgE and fragments of IgE have been analyzed using surface plasmon resonance. As 
      described in the preceding paper of this issue [Henry, A. J., et al. (1997) 
      Biochemistry 36, 15568-15578], biphasic binding kinetics was observed. Two of the 
      mutations had significant effects on binding: K117D reduced the affinity of 
      sFcepsilonRIalpha for IgE by a factor of 30, while D159K increased the affinity 
      for IgE by a factor of 7, both principally through changes in the rates of 
      dissociation of the slower phase of the interaction. Circular dichroism spectra 
      of sFcepsilonRIalpha incorporating either of these mutations were 
      indistinguishable from those of wild-type sFcepsilonRIalpha, demonstrating that 
      the native conformation had not been disrupted. Our results, together with those 
      from site-directed mutagenesis on fragments of IgE presented in the accompanying 
      paper, define the contact surfaces in the IgE:sFcepsilonRIalpha complex.
FAU - Cook, J P
AU  - Cook JP
AD  - The Randall Institute, King's College London, 26-29 Drury Lane, London, WC2B 5RL, 
      United Kingdom.
FAU - Henry, A J
AU  - Henry AJ
FAU - McDonnell, J M
AU  - McDonnell JM
FAU - Owens, R J
AU  - Owens RJ
FAU - Sutton, B J
AU  - Sutton BJ
FAU - Gould, H J
AU  - Gould HJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Immunoglobulin Fc Fragments)
RN  - 0 (Receptors, IgE)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Binding Sites
MH  - Biosensing Techniques
MH  - Circular Dichroism
MH  - Cloning, Molecular
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Humans
MH  - Hypersensitivity/etiology
MH  - Immunoglobulin E/chemistry/*metabolism
MH  - Immunoglobulin Fc Fragments/chemistry/*metabolism
MH  - Kinetics
MH  - Models, Molecular
MH  - Mutagenesis, Site-Directed
MH  - Protein Binding
MH  - Protein Conformation
MH  - Receptors, IgE/*chemistry/genetics/*metabolism
MH  - Transfection/genetics
MH  - Tumor Cells, Cultured
EDAT- 1998/01/31 00:00
MHDA- 1998/01/31 00:01
CRDT- 1998/01/31 00:00
PHST- 1998/01/31 00:00 [pubmed]
PHST- 1998/01/31 00:01 [medline]
PHST- 1998/01/31 00:00 [entrez]
AID - bi9713005 [pii]
AID - 10.1021/bi9713005 [doi]
PST - ppublish
SO  - Biochemistry. 1997 Dec 16;36(50):15579-88. doi: 10.1021/bi9713005.