Linked Life Data

11897784

Source:http://linkedlifedata.com/resource/pubmed/id/11897784

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
21
pubmed:dateCreated
2002-5-20
pubmed:abstractText
In response to accumulation of unfolded proteins in the endoplasmic reticulum (ER), cells activate an intracellular signal transduction pathway called the unfolded protein response (UPR). IRE and PERK are the two type-I ER transmembrane protein kinase receptors that signal the UPR. The N-terminal luminal domains (NLDs) of IRE1 and PERK sense ER stress conditions by a common mechanism and transmit the signal to regulate the cytoplasmic domains of these receptors. To provide an experimental system amenable to detailed biochemical and structural analysis to elucidate the mechanism of ER-transmembrane signaling mechanism mediated by the NLD, we overexpressed the soluble luminal domain of human IRE1alpha in COS-1 cells by transient DNA transfection. Here we report the expression, purification, and characterization of the soluble NLD. The biological function of the NLD was confirmed by its ability to associate with itself and to interact with both the membrane-bound full-length IRE1alpha receptor and the ER chaperone BiP. Functional and spectral studies suggested that the highly conserved N-linked glycosylation site is not required for proper protein folding and self-association. Interestingly, we demonstrated that the NLD forms stable dimers linked by intermolecular disulfide bridges. Our data support that the luminal domain represents a novel ligand-independent dimerization domain.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
24
pubmed:volume
277
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
18346-56
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:11897784-Amino Acid Sequence, pubmed-meshheading:11897784-Animals, pubmed-meshheading:11897784-COS Cells, pubmed-meshheading:11897784-Circular Dichroism, pubmed-meshheading:11897784-Dimerization, pubmed-meshheading:11897784-Endoribonucleases, pubmed-meshheading:11897784-Fungal Proteins, pubmed-meshheading:11897784-Humans, pubmed-meshheading:11897784-Ligands, pubmed-meshheading:11897784-Membrane Glycoproteins, pubmed-meshheading:11897784-Membrane Proteins, pubmed-meshheading:11897784-Molecular Sequence Data, pubmed-meshheading:11897784-Protein Denaturation, pubmed-meshheading:11897784-Protein-Serine-Threonine Kinases, pubmed-meshheading:11897784-Saccharomyces cerevisiae Proteins, pubmed-meshheading:11897784-Spectrometry, Fluorescence, pubmed-meshheading:11897784-Spectrometry, Mass, Matrix-Assisted Laser..., pubmed-meshheading:11897784-Transfection
pubmed:year
2002
pubmed:articleTitle
The protein kinase/endoribonuclease IRE1alpha that signals the unfolded protein response has a luminal N-terminal ligand-independent dimerization domain.
pubmed:affiliation
Howard Hughes Medical Institute, Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan 48109-0650, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't