Linked Life Data

16519528

Source:http://linkedlifedata.com/resource/pubmed/id/16519528

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2006-3-7
pubmed:abstractText
RPE65 is the major component of the retinal pigment epithelium (RPE) microsomal membrane, and it plays a critical role in the binding of retinoids involved in the visual cycle. To understand how RPE65 binds to membranes, we have expressed and purified soluble fragments of human RPE65 fused to glutathione S-transferase (GST). The interaction between two fragments of RPE65 (F1 and F2 which include residues 1-125 and 126-250, respectively) and lipid monolayers has been studied by surface pressure, ellipsometry, and surface rheology measurements. Surface pressure and ellipsometry clearly showed a rapid adsorption of F2 to lipid monolayers whereas the kinetics of binding of F1 was much slower. Furthermore, the data suggest that the F2 fragment inserts into the lipid monolayer. Surface rheology showed a clear increase in monolayer rigidity only in the presence of F2, thereby demonstrating high intermolecular interactions of this fragment. This observation is further supported by the GST pull-down assays which demonstrated that F2 cosediments with full-length RPE65, suggesting that RPE65 has the propensity to form clusters or oligomers. The structure homology modeling of RPE65 based on a related family member, apocarotene 15',15'-oxygenase, further suggests that a hydrophobic patch located in the F2 region might be responsible for membrane binding. The present work shows that F2 interacts much stronger with lipid monolayers than does F1, which suggests that the region of RPE65 located between residues 126-250 should be very important for its membrane binding. Moreover, given that these fragments are not acylated, these data also suggest that an effective binding of RPE65 to membranes can be achieved without palmitoylation. Furthermore, GST pull-down assays also indicated that F2 interacts with 11-cis-retinol dehydrogenase, which supports previous data suggesting that it could act as a partner of RPE65.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
14
pubmed:volume
45
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3337-47
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed-meshheading:16519528-Amino Acid Sequence, pubmed-meshheading:16519528-Animals, pubmed-meshheading:16519528-Baculoviridae, pubmed-meshheading:16519528-Carrier Proteins, pubmed-meshheading:16519528-Cell Membrane, pubmed-meshheading:16519528-Escherichia coli, pubmed-meshheading:16519528-Eye Proteins, pubmed-meshheading:16519528-Glutathione Transferase, pubmed-meshheading:16519528-Humans, pubmed-meshheading:16519528-Hydrophobic and Hydrophilic Interactions, pubmed-meshheading:16519528-Insects, pubmed-meshheading:16519528-Molecular Sequence Data, pubmed-meshheading:16519528-Oxidoreductases, pubmed-meshheading:16519528-Oxygenases, pubmed-meshheading:16519528-Protein Binding, pubmed-meshheading:16519528-Protein Structure, Tertiary, pubmed-meshheading:16519528-Recombinant Fusion Proteins, pubmed-meshheading:16519528-Sequence Alignment, pubmed-meshheading:16519528-Solubility, pubmed-meshheading:16519528-Time Factors, pubmed-meshheading:16519528-beta-Carotene 15,15'-Monooxygenase
pubmed:year
2006
pubmed:articleTitle
Binding of RPE65 fragments to lipid monolayers and identification of its partners by glutathione S-transferase pull-down assays.
pubmed:affiliation
Unité de Recherche en Ophtalmologie, Université Laval, Québec, Québec, Canada G1V 4G2.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't