Source:http://linkedlifedata.com/resource/pubmed/id/12899628
Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
31
|
| pubmed:dateCreated |
2003-8-5
|
| pubmed:abstractText |
To investigate structure and function relations of a new member of the exchangeable apolipoprotein family that modulates plasma lipid levels, recombinant human apolipoprotein (apo) A-V was produced in Escherichia coli and isolated by a combination of nickel chelation affinity chromatography and reversed-phase HPLC. Antibodies directed against apoA-V were generated and employed in immunoblotting experiments. Anti-apoA-V IgG gave a strong response against recombinant apoA-V from E. coli and human apoA-V expressed in transgenic mice, but did not recognize human apoA-I or apoA-IV. In neutral-pH buffers, at concentrations of >0.1 mg/mL, isolated lipid-free apoA-V is poorly soluble. By contrast, apoA-V is soluble in 50 mM sodium citrate (pH 3.0). Far-UV circular dichroism analysis and spectral deconvolution reveal that apoA-V possesses 32% alpha-helix, 33% beta-sheet, 16% beta-turn, and 18% random coil secondary structure conformers. Temperature-induced denaturation studies gave rise to a transition midpoint of 47.1 degrees C. Upon being cooled to ambient temperature from 85 degrees C, apoA-V failed to recover all of the negative ellipticity present in unheated apoA-V. ApoA-V interacts with bilayer vesicles of dimyristoylphosphatidylcholine to form discoidal complexes with diameters in the range of 15-20 nm. However, apoA-V was a poor activator of lecithin:cholesterol acyltransferase where the activity was 8.5 +/- 1.8% of that of apoA-I. Furthermore, apoA-V failed to support enhanced efflux of cholesterol from cAMP-treated J774 macrophages, although low levels of efflux were obtained from unstimulated cells. Taken together, the results demonstrate recombinant apoA-V possesses unique structural and functional characteristics, in keeping with its proposed role in the modulation of plasma lipid levels.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/APOA5 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Apoa5 protein, mouse,
http://linkedlifedata.com/resource/pubmed/chemical/Apolipoprotein A-I,
http://linkedlifedata.com/resource/pubmed/chemical/Apolipoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Apolipoproteins A,
http://linkedlifedata.com/resource/pubmed/chemical/Cholesterol,
http://linkedlifedata.com/resource/pubmed/chemical/Dimyristoylphosphatidylcholine,
http://linkedlifedata.com/resource/pubmed/chemical/Glutathione Transferase,
http://linkedlifedata.com/resource/pubmed/chemical/Lipid Bilayers,
http://linkedlifedata.com/resource/pubmed/chemical/Lipids,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphatidylcholine-Sterol...,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/apolipoprotein A-IV
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Aug
|
| pubmed:issn |
0006-2960
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
12
|
| pubmed:volume |
42
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
9416-23
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:12899628-Animals,
pubmed-meshheading:12899628-Apolipoprotein A-I,
pubmed-meshheading:12899628-Apolipoproteins,
pubmed-meshheading:12899628-Apolipoproteins A,
pubmed-meshheading:12899628-Cholesterol,
pubmed-meshheading:12899628-Circular Dichroism,
pubmed-meshheading:12899628-Dimyristoylphosphatidylcholine,
pubmed-meshheading:12899628-Escherichia coli,
pubmed-meshheading:12899628-Gene Expression,
pubmed-meshheading:12899628-Glutathione Transferase,
pubmed-meshheading:12899628-Goats,
pubmed-meshheading:12899628-Homeostasis,
pubmed-meshheading:12899628-Humans,
pubmed-meshheading:12899628-Immunoblotting,
pubmed-meshheading:12899628-Lipid Bilayers,
pubmed-meshheading:12899628-Lipids,
pubmed-meshheading:12899628-Mice,
pubmed-meshheading:12899628-Mice, Transgenic,
pubmed-meshheading:12899628-Phosphatidylcholine-Sterol O-Acyltransferase,
pubmed-meshheading:12899628-Protein Binding,
pubmed-meshheading:12899628-Protein Conformation,
pubmed-meshheading:12899628-Recombinant Proteins,
pubmed-meshheading:12899628-Structure-Activity Relationship
|
| pubmed:year |
2003
|
| pubmed:articleTitle |
Structure-function studies of human apolipoprotein A-V: a regulator of plasma lipid homeostasis.
|
| pubmed:affiliation |
Lipid Biology in Health and Disease Research Group, Children's Hospital Oakland Research Institute, Oakland, California 94609, USA.
|
| pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|