Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
|
| pubmed:dateCreated |
1994-7-28
|
| pubmed:abstractText |
The GS protein alpha subunit (alpha S) sequences conserved in non-myristoylated alpha subunits (TENIR, residues 369-373) or critical for adenylyl cyclase interaction were investigated as possible sites required for membrane localization. Substitutions were created by site-directed mutagenesis in which the TENIR residues were deleted from alpha S or added to the soluble, non-myristoylated alpha i1. After transfection, COS cells were separated by centrifugation into particulate and soluble fractions. Immunoblots showed that these substitutions did not change the localization: alpha S +/- TENIR in the particulate fraction, non-myristoylated alpha i1 +/- TENIR in the soluble fraction. The constitutively active alpha i/alpha S chimera (CH4A), containing four regions of alpha S sufficient for adenylyl cyclase activation, was mutated to prevent myristoylation (GA-CH4A). Immunoblots of transfected COS cell fractions showed CH4A in the particulate and GA-CH4A in the soluble fraction. While these regions did not lead to membrane localization, the soluble GA-CH4A could activate adenylyl cyclase in the intact cell and after reconstitution with cyc- membranes.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jan
|
| pubmed:issn |
0898-6568
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
6
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
25-33
|
| pubmed:dateRevised |
2005-11-17
|
| pubmed:meshHeading |
pubmed-meshheading:8011426-Adenylate Cyclase,
pubmed-meshheading:8011426-Amino Acid Sequence,
pubmed-meshheading:8011426-Animals,
pubmed-meshheading:8011426-Base Sequence,
pubmed-meshheading:8011426-Cell Line, Transformed,
pubmed-meshheading:8011426-Cell Membrane,
pubmed-meshheading:8011426-Cells, Cultured,
pubmed-meshheading:8011426-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8011426-GTP-Binding Proteins,
pubmed-meshheading:8011426-Gene Expression,
pubmed-meshheading:8011426-Macaca,
pubmed-meshheading:8011426-Molecular Sequence Data,
pubmed-meshheading:8011426-Mutagenesis, Site-Directed,
pubmed-meshheading:8011426-Oligopeptides,
pubmed-meshheading:8011426-Recombinant Fusion Proteins,
pubmed-meshheading:8011426-Structure-Activity Relationship,
pubmed-meshheading:8011426-Transfection
|
| pubmed:year |
1994
|
| pubmed:articleTitle |
The membrane localization of the G protein alpha s subunit is not dependent on its TENIR sequence or effector domain.
|
| pubmed:affiliation |
Molecular Pathophysiology Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.
|
| pubmed:publicationType |
Journal Article
|