Linked Life Data

10504303

Source:http://linkedlifedata.com/resource/pubmed/id/10504303

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2000-1-10
pubmed:abstractText
We have investigated recombinant fibrillin-1 (profib-1) and fibrillin-2 (glyfib-2) molecules encoding the proline- or glycine-rich regions with flanking domains (exons 9-11), in order to establish whether these sequences might mediate specific molecular recognition events important in fibrillin assembly. Our data demonstrate that both recombinant molecules can form extracellular dimers, but highlight subtle differences in the stability of these dimers. Following expression in COS-1 cells, SDS-PAGE analysis showed that glyfib-2 was present intracellularly as monomers, and extracellularly as monomers and disulphide-bonded dimers. Size fractionation in native non-reducing conditions prior to SDS-PAGE analysis highlighted that glyfib-2 also formed non-covalent associations. In contrast, profib-1 appeared monomeric in cells and medium. Using an in vitro translation system supplemented with semipermeabilised HT1080 cells together with chemical crosslinking, dimers of the fibrillin-1 and fibrillin-2 molecules were detected. Dimerisation was not cell-dependent since molecules translated in the absence of cells dimerised, and was not an intracellular event as judged by proteinase K digestions. A crosslinking and coimmunoprecipitation strategy provided a means of investigating whether molecular chaperones might be involved in preventing dimerisation of translocated molecules. Proteinase K-resistant recombinant molecules associated rapidly with BiP, and thereafter with protein disulphide isomerase and calreticulin. Differences between the two fibrillin isoforms in ability to form stable dimers prompted investigation of the proline- and glycine-rich sequences. Differences in solubility and pI were apparent that may contribute to reduced stability of proline-rich region interactions. These studies suggest that extracellular dimer formation mediated by interactions of the proline- and glycine-rich regions may be a crucial early step in the extracellular assembly of fibrillin into microfibrils.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0021-9533
pubmed:author
pubmed:issnType
Print
pubmed:volume
112 ( Pt 20)
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3549-58
pubmed:dateRevised
2009-9-29
pubmed:meshHeading
pubmed-meshheading:10504303-Amino Acid Sequence, pubmed-meshheading:10504303-Animals, pubmed-meshheading:10504303-COS Cells, pubmed-meshheading:10504303-Calcium-Binding Proteins, pubmed-meshheading:10504303-Calnexin, pubmed-meshheading:10504303-Calreticulin, pubmed-meshheading:10504303-Cell Line, pubmed-meshheading:10504303-Dimerization, pubmed-meshheading:10504303-Exons, pubmed-meshheading:10504303-Extracellular Matrix Proteins, pubmed-meshheading:10504303-Glycine, pubmed-meshheading:10504303-Humans, pubmed-meshheading:10504303-Microfilament Proteins, pubmed-meshheading:10504303-Proline, pubmed-meshheading:10504303-Protein Biosynthesis, pubmed-meshheading:10504303-Recombinant Proteins, pubmed-meshheading:10504303-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:10504303-Ribonucleoproteins, pubmed-meshheading:10504303-Transcription, Genetic, pubmed-meshheading:10504303-Transfection
pubmed:year
1999
pubmed:articleTitle
Fibrillin assembly: dimer formation mediated by amino-terminal sequences.
pubmed:affiliation
Wellcome Trust Centre for Cell-Matrix Research, School of Biological Sciences, University of Manchester, Oxford Road, Manchester, M13 9PT, UK. cay.kielty@man.ac.uk
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't