Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
13
pubmed:dateCreated
2000-5-4
pubmed:abstractText
The murine class B, type I scavenger receptor (mSR-BI) is a receptor for both high density lipoprotein (HDL) and low density lipoprotein (LDL) and mediates selective, rather than endocytic, uptake of lipoprotein lipid. We have developed a "retrovirus library-based activity dissection" method to generate mSR-BI mutants in which some, but not all, of the activities of this multifunctional protein have been disrupted. This method employs three techniques: 1) efficient in vitro cDNA mutagenesis (here error-prone PCR was used), 2) efficient retroviral delivery and high expression of single mutant cDNAs into individual cells, and 3) isolation of infected cells expressing the desired mutant phenotype using high sensitivity positive/negative screening by two-color fluorescence-activated cell sorting. A set of mutants, all having arginine substitutions at two common sites (positions 402 or 401 and position 418), were isolated and characterized. Mutation at either site alone did not generate as strong a mutant phenotype (loss of DiI uptake from DiI-HDL) as did the double mutations. "Activity-dissected" double mutants were as effective as wild-type mSR-BI in functioning as LDL receptors, mediating high affinity LDL binding and uptake of metabolically active cholesterol from LDL, but they lost most of their corresponding HDL receptor activity. Thus, these mutants provide support for the proposal that the interaction of SR-BI with HDL differs from that with LDL. Examination of the in vivo function of such mutants may provide insights into the differential roles of the LDL and HDL receptor activities of SR-BI in normal lipoprotein metabolism and in SR-BI's ability to protect against atherosclerosis.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, CD36, http://linkedlifedata.com/resource/pubmed/chemical/Arginine, http://linkedlifedata.com/resource/pubmed/chemical/Cholesterol Esters, http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers, http://linkedlifedata.com/resource/pubmed/chemical/Lipoproteins, HDL, http://linkedlifedata.com/resource/pubmed/chemical/Lipoproteins, LDL, http://linkedlifedata.com/resource/pubmed/chemical/Membrane Proteins, http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Immunologic, http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Lipoprotein, http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Scavenger, http://linkedlifedata.com/resource/pubmed/chemical/Scarb1 protein, mouse, http://linkedlifedata.com/resource/pubmed/chemical/Scavenger Receptors, Class B
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
31
pubmed:volume
275
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
9120-30
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:10734045-Amino Acid Sequence, pubmed-meshheading:10734045-Animals, pubmed-meshheading:10734045-Antigens, CD36, pubmed-meshheading:10734045-Arginine, pubmed-meshheading:10734045-Base Sequence, pubmed-meshheading:10734045-CHO Cells, pubmed-meshheading:10734045-Cholesterol Esters, pubmed-meshheading:10734045-Cloning, Molecular, pubmed-meshheading:10734045-Cricetinae, pubmed-meshheading:10734045-DNA Primers, pubmed-meshheading:10734045-Genomic Library, pubmed-meshheading:10734045-Lipoproteins, HDL, pubmed-meshheading:10734045-Lipoproteins, LDL, pubmed-meshheading:10734045-Membrane Proteins, pubmed-meshheading:10734045-Mice, pubmed-meshheading:10734045-Molecular Sequence Data, pubmed-meshheading:10734045-Mutagenesis, pubmed-meshheading:10734045-Receptors, Immunologic, pubmed-meshheading:10734045-Receptors, Lipoprotein, pubmed-meshheading:10734045-Receptors, Scavenger, pubmed-meshheading:10734045-Retroviridae, pubmed-meshheading:10734045-Scavenger Receptors, Class B
pubmed:year
2000
pubmed:articleTitle
Dissociation of the high density lipoprotein and low density lipoprotein binding activities of murine scavenger receptor class B type I (mSR-BI) using retrovirus library-based activity dissection.
pubmed:affiliation
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't