PMID- 8886836
OWN - NLM
STAT- MEDLINE
DCOM- 19970403
LR  - 20131121
IS  - 0956-960X (Print)
IS  - 0956-960X (Linking)
VI  - 7
IP  - 1
DP  - 1996
TI  - Unsusceptibility of recombinant human Fc fragments of immunoglobulin E to 
      thrombin.
PG  - 42-7
AB  - Human immunoglobulin E (IgE) contains a potential recognition sequence for 
      thrombin protease cleavage at N-terminal end of C epsilon 3 domain responsible 
      for binding to alpha chain of high affinity Fc receptor for IgE (Fc epsilon RI 
      alpha), but it remains unknown for the enzyme susceptibility. The human Fc 
      fragments of IgE (IgE-Fc), consisting of C epsilon 2' (Ala329)-C epsilon 3-C 
      epsilon 4 chains (Fc') and of C epsilon 2' (Thr315-Ala329)-C epsilon 3-C epsilon 
      4 chains (F(c')2), were expressed in the mammalian COS and Chinese hamster ovary 
      (CHO) cells by placing the IgE-Fc cDNA under the control of the cytomegalovirus 
      (CMV) promoter. Under nonreducing condition F(c')2 was not cleaved by thrombin 
      protease as well as native IgE. Neither treatment of F(c')2 with final 0.1% 
      2-mercaptoethanol at boiling point for 5 min, with a sulfhydryl-reactive biotin 
      derivative, by which monomers in COS-derived F(c')2 preparations were 
      biotinylated at Cys328, nor with neuraminidase, affected the accessibility to the 
      enzyme. These results suggested that F(c')2, either dimeric or monomeric, had 
      compact conformations.
FAU - Kamiya, T
AU  - Kamiya T
AD  - Central Research Laboratories, Green Cross Corporation, Osaka, Japan.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Hum Antibodies Hybridomas
JT  - Human antibodies and hybridomas
JID - 9014461
RN  - 0 (Immunoglobulin Fc Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - 60-24-2 (Mercaptoethanol)
RN  - 6SO6U10H04 (Biotin)
RN  - EC 3.2.1.18 (Neuraminidase)
RN  - EC 3.4.21.5 (Thrombin)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - Biotin/analogs & derivatives
MH  - CHO Cells
MH  - COS Cells
MH  - Cricetinae
MH  - Humans
MH  - Immunoglobulin E/*chemistry
MH  - Immunoglobulin Fc Fragments/biosynthesis/*chemistry/genetics
MH  - Mercaptoethanol
MH  - Neuraminidase
MH  - Protein Conformation
MH  - Recombinant Proteins/chemistry
MH  - Thrombin/chemistry/*metabolism
MH  - Transfection
EDAT- 1996/01/01 00:00
MHDA- 1996/01/01 00:01
CRDT- 1996/01/01 00:00
PHST- 1996/01/01 00:00 [pubmed]
PHST- 1996/01/01 00:01 [medline]
PHST- 1996/01/01 00:00 [entrez]
PST - ppublish
SO  - Hum Antibodies Hybridomas. 1996;7(1):42-7.