PMID- 10486146
OWN - NLM
STAT- MEDLINE
DCOM- 19991103
LR  - 20071115
IS  - 0003-9861 (Print)
IS  - 0003-9861 (Linking)
VI  - 369
IP  - 2
DP  - 1999 Sep 15
TI  - Characterization of recombinant human CXCR4 in insect cells: role of
      extracellular domains and N-glycosylation in ligand binding.
PG  - 267-76
AB  - The cDNA of the human CXCR4/fusin was isolated from a human HeLa cell cDNA
      library by PCR and functionally expressed in Sf9 insect cells. The recombinant
      receptor was found to bind its natural ligand SDF-1alpha with an affinity
      comparable to that of the native receptor. Sequence-specific antibodies against
      each of the four extracellular domains were generated and used to investigate the
      interactions between the different domains of the receptor and the ligand. Each
      of the four antibodies was found to be able to inhibit ligand binding. CXCR4 was 
      shown to be a glycoprotein. The role of N-glycosylation of CXCR4 in ligand
      binding was investigated in the insect cells overexpressed with recombinant
      CXCR4. Two potential N-glycosylation sites (Asn-11 and Asn-176) were either
      singly or doubly mutated to a leucine residue. Both single mutant receptors
      exhibited a significant decrease in ligand binding activity and affinity. The
      double mutant receptor showed little binding activity. Our data suggest that all 
      of the extracellular domains are involved in ligand-receptor interactions and
      that N-glycosylation is required to maintain high-affinity ligand binding.
FAU - Zhou, H
AU  - Zhou H
AD  - Division of Pharmaceutical Sciences, College of Pharmacy, Lexington, Kentucky,
      40536-0082, USA.
FAU - Tai, H H
AU  - Tai HH
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Arch Biochem Biophys
JT  - Archives of biochemistry and biophysics
JID - 0372430
RN  - 0 (CXCL12 protein, human)
RN  - 0 (Chemokine CXCL12)
RN  - 0 (Chemokines, CXC)
RN  - 0 (DNA, Complementary)
RN  - 0 (Ligands)
RN  - 0 (Peptide Fragments)
RN  - 0 (Receptors, CXCR4)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Animals
MH  - Antibody Specificity
MH  - Baculoviridae
MH  - Chemokine CXCL12
MH  - Chemokines, CXC/*metabolism
MH  - DNA, Complementary
MH  - Gene Library
MH  - Glycosylation
MH  - HeLa Cells
MH  - Humans
MH  - Ligands
MH  - Peptide Fragments/genetics/immunology/isolation & purification/*metabolism
MH  - Protein Processing, Post-Translational
MH  - Receptors, CXCR4/genetics/immunology/isolation & purification/*metabolism
MH  - Recombinant Proteins/immunology/isolation & purification/metabolism
MH  - Spodoptera
EDAT- 1999/09/16 00:00
MHDA- 1999/09/16 00:01
CRDT- 1999/09/16 00:00
PHST- 1999/09/16 00:00 [pubmed]
PHST- 1999/09/16 00:01 [medline]
PHST- 1999/09/16 00:00 [entrez]
AID - 10.1006/abbi.1999.1368 [doi]
AID - S0003-9861(99)91368-1 [pii]
PST - ppublish
SO  - Arch Biochem Biophys. 1999 Sep 15;369(2):267-76. doi: 10.1006/abbi.1999.1368.