Linked Life Data

15100222

Source:http://linkedlifedata.com/resource/pubmed/id/15100222

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
26
pubmed:dateCreated
2004-6-21
pubmed:abstractText
Cellular retinaldehyde-binding protein (CRALBP) functions in the retinal pigment epithelium (RPE) as an acceptor of 11-cis-retinol in the isomerization step of the rod visual cycle and as a substrate carrier for 11-cis-retinol dehydrogenase. Toward a better understanding of CRALBP function, the ligand binding cavity in human recombinant CRALBP (rCRALBP) was characterized by photoaffinity labeling with 3-diazo-4-keto-11-cis-retinal and by high resolution mass spectrometric topological analyses. Eight photoaffinity-modified residues were identified in rCRALBP by liquid chromatography tandem mass spectrometry, including Tyr(179), Phe(197), Cys(198), Met(208), Lys(221), Met(222), Val(223), and Met(225). Multiple different adduct masses were found on the photolabeled residues, and the molecular identity of each modification remains unknown. Supporting the specificity of photo-labeling, 50% of the modified residues have been associate with retinoid interactions by independent analyses. In addition, topological analysis of apo- and holo-rCRALBP by hydrogen/deuterium exchange and mass spectrometry demonstrated residues 198-255 incorporate significantly less deuterium when the retinoid binding pocket is occupied with 11-cis-retinal. This hydrophobic region encompasses all but one of the photo-labeled residues. A structural model of CRALBP ligand binding domain was constructed based on the crystal structures of three homologues in the CRAL-TRIO family of lipid-binding proteins. In the model, all of the photolabeled residues line the ligand binding cavity except Met(208), which appears to reside in a flexible loop at the entrance/exit of the ligand cavity. Overall, the results expand to 12 the number of residues proposed to interact with ligand and provide further insight into CRALBP ligand and protein interactions.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
279
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
27357-64
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:15100222-Amino Acid Sequence, pubmed-meshheading:15100222-Carrier Proteins, pubmed-meshheading:15100222-Deuterium Exchange Measurement, pubmed-meshheading:15100222-Escherichia coli, pubmed-meshheading:15100222-Humans, pubmed-meshheading:15100222-Hydrogen, pubmed-meshheading:15100222-Isomerism, pubmed-meshheading:15100222-Ligands, pubmed-meshheading:15100222-Models, Molecular, pubmed-meshheading:15100222-Molecular Sequence Data, pubmed-meshheading:15100222-Peptide Fragments, pubmed-meshheading:15100222-Photoaffinity Labels, pubmed-meshheading:15100222-Protein Binding, pubmed-meshheading:15100222-Protein Structure, Tertiary, pubmed-meshheading:15100222-Recombinant Proteins, pubmed-meshheading:15100222-Retinaldehyde, pubmed-meshheading:15100222-Retinoids, pubmed-meshheading:15100222-Spectrometry, Mass, Electrospray Ionization, pubmed-meshheading:15100222-Spectrophotometry, Ultraviolet, pubmed-meshheading:15100222-Tritium
pubmed:year
2004
pubmed:articleTitle
Identification of CRALBP ligand interactions by photoaffinity labeling, hydrogen/deuterium exchange, and structural modeling.
pubmed:affiliation
Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't