PMID- 14680941
OWN - NLM
STAT- MEDLINE
DCOM- 20040630
LR  - 20131121
IS  - 1046-5928 (Print)
IS  - 1046-5928 (Linking)
VI  - 32
IP  - 1
DP  - 2003 Nov
TI  - Refolding and characterization of the functional ligand-binding domain of human 
      lectin-like oxidized LDL receptor.
PG  - 68-74
AB  - Lectin-like oxidized low-density lipoprotein receptor (LOX-1), a type II membrane 
      protein that can recognize a variety of structurally unrelated macromolecules, 
      plays an important role in host defense and is implicated in atherogenesis. To 
      understand the interaction between human LOX-1 and its ligands, in this study the 
      functional C-type lectin-like domain (CTLD) of LOX-1 was reconstituted at high 
      efficiency from inactive aggregates in Escherichia coli using a refolding 
      technique based on an artificial chaperone. The CD spectra of the purified domain 
      suggested that the domain has alpha-helical structure and the blue shift of Trp 
      residues was observed on refolding of the domain. Like wild-type hLOX-1, the 
      refolded CTLD domain was able to bind modified LDL. Thus, even though CTLD 
      contains six Cys residues that form disulfide bonds, it recovered its specific 
      binding ability on refolding. This suggests that the correct disulfide bonds in 
      CTLD were formed by the artificial chaperone technique. Although the domain 
      lacked N-glycosylation, it showed high affinity for its ligand in surface plasmon 
      resonance experiments. Thus, unglycosylated CTLD is sufficient for binding 
      modified LDL.
FAU - Xie, Qiuhong
AU  - Xie Q
AD  - National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, 
      Japan.
FAU - Matsunaga, Shigeru
AU  - Matsunaga S
FAU - Shi, Xiaohua
AU  - Shi X
FAU - Ogawa, Setsuko
AU  - Ogawa S
FAU - Niimi, Setsuko
AU  - Niimi S
FAU - Wen, Zhesheng
AU  - Wen Z
FAU - Tokuyasu, Ken
AU  - Tokuyasu K
FAU - Machida, Sachiko
AU  - Machida S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (Ligands)
RN  - 0 (OLR1 protein, human)
RN  - 0 (Receptors, LDL)
RN  - 0 (Receptors, Oxidized LDL)
RN  - 0 (Scavenger Receptors, Class E)
RN  - 8DUH1N11BX (Tryptophan)
SB  - IM
MH  - Binding Sites
MH  - Circular Dichroism
MH  - Escherichia coli
MH  - Glycosylation
MH  - Humans
MH  - Ligands
MH  - *Protein Folding
MH  - Protein Renaturation
MH  - Protein Structure, Tertiary
MH  - Receptors, LDL/*chemistry/genetics/*metabolism
MH  - Receptors, Oxidized LDL
MH  - Scavenger Receptors, Class E
MH  - Spectrometry, Fluorescence
MH  - Surface Plasmon Resonance
MH  - Tryptophan/chemistry
EDAT- 2003/12/19 05:00
MHDA- 2004/07/01 05:00
CRDT- 2003/12/19 05:00
PHST- 2003/04/04 00:00 [received]
PHST- 2003/07/08 00:00 [revised]
PHST- 2003/12/19 05:00 [pubmed]
PHST- 2004/07/01 05:00 [medline]
PHST- 2003/12/19 05:00 [entrez]
AID - S1046592803002201 [pii]
AID - 10.1016/S1046-5928(03)00220-1 [doi]
PST - ppublish
SO  - Protein Expr Purif. 2003 Nov;32(1):68-74. doi: 10.1016/S1046-5928(03)00220-1.