Linked Life Data

12110670

Source:http://linkedlifedata.com/resource/pubmed/id/12110670

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
39
pubmed:dateCreated
2002-9-23
pubmed:abstractText
We have investigated the folding of the myosin motor domain using a chimera of an embryonic striated muscle myosin II motor domain fused on its COOH terminus to a thermal stable, fast folding variant of green fluorescent protein (GFP). In in vitro expression assays, the GFP domain of the chimeric protein, S1(795)GFP, folds rapidly enabling us to monitor the folding of the motor domain using fluorescence. The myosin motor domain folds very slowly and transits through multiple intermediates that are detectable by gel filtration chromatography. The distribution of the nascent protein among these intermediates is strongly dependent upon temperature. At 25 degrees C and above the predominant product is an aggregate of S1(795)GFP or a complex with other lysate proteins. At 0 degrees C, the motor domain folds slowly via an energy independent pathway. The unusual temperature dependence and slow rate suggests that folding of the myosin motor is highly susceptible to off-pathway interactions and aggregation. Expression of the S1(795)GFP in the C2C12 muscle cell line yields a folded and functionally active protein that exhibits Mg(2+)ATP-sensitive actin-binding and myosin motor activity. In contrast, expression of S1(795)GFP in kidney epithelial cell lines (human 293 and COS 7 cells) results in an inactive and aggregated protein. The results of the in vitro folding assay suggest that the myosin motor domain does not fold spontaneously under physiological conditions and probably requires cytosolic chaperones. The expression studies support this conclusion and demonstrate that these factors are optimized in muscle cells.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
27
pubmed:volume
277
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
36799-807
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:12110670-Actins, pubmed-meshheading:12110670-Adenosine Triphosphate, pubmed-meshheading:12110670-Adenoviridae, pubmed-meshheading:12110670-Ammonium Sulfate, pubmed-meshheading:12110670-Animals, pubmed-meshheading:12110670-COS Cells, pubmed-meshheading:12110670-Cell Line, pubmed-meshheading:12110670-Chickens, pubmed-meshheading:12110670-Chromatography, Gel, pubmed-meshheading:12110670-Cytoplasm, pubmed-meshheading:12110670-DNA, Complementary, pubmed-meshheading:12110670-Densitometry, pubmed-meshheading:12110670-Green Fluorescent Proteins, pubmed-meshheading:12110670-Humans, pubmed-meshheading:12110670-Luminescent Proteins, pubmed-meshheading:12110670-Microscopy, Fluorescence, pubmed-meshheading:12110670-Models, Molecular, pubmed-meshheading:12110670-Muscle, Skeletal, pubmed-meshheading:12110670-Muscles, pubmed-meshheading:12110670-Myosins, pubmed-meshheading:12110670-Protein Binding, pubmed-meshheading:12110670-Protein Biosynthesis, pubmed-meshheading:12110670-Protein Folding, pubmed-meshheading:12110670-Protein Structure, Tertiary, pubmed-meshheading:12110670-Recombinant Fusion Proteins, pubmed-meshheading:12110670-Temperature, pubmed-meshheading:12110670-Transcription, Genetic
pubmed:year
2002
pubmed:articleTitle
Folding of the striated muscle myosin motor domain.
pubmed:affiliation
Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, New Jersey 08854, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't