Linked Life Data

11257508

Source:http://linkedlifedata.com/resource/pubmed/id/11257508

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2001-3-21
pubmed:abstractText
C-Terminal binding protein (CtBP) interacts with a highly conserved amino acid motif (PXDLS) at the C terminus of adenovirus early region 1A (AdE1A) protein. This amino acid sequence has recently been demonstrated in the mammalian protein C-terminal interacting protein (CtIP) and a number of Drosophila repressors including Snail, Knirps and Hairy. In the study described here we have examined the structures of synthetic peptides identical to the CtBP binding sites on these proteins using NMR spectroscopy. It has been shown that peptides identical to the CtBP binding site in CtIP and at the N terminus of Snail form a series of beta-turns similar to those seen in AdE1A. The PXDLS motif towards the C terminus of Snail forms an alpha-helix. However, the motifs in Knirps and Hairy did not adopt well-defined structures in TFE/water mixtures as shown by the absence of medium range NOEs and a high proportion of signal overlap. The affinities of peptides for Drosophila and mammalian CtBP were compared using enzyme-linked immunosorbent assay. CtIP, Snail (N-terminal peptide) and Knirps peptides all bind to mammalian CtBP with high affinity (K(i) of 1.04, 1.34 and 0.52 microM, respectively). However, different effects were observed with dCtBP, most notably the affinity for the Snail (N-terminal peptide) and Knirps peptides were markedly reduced (K(i) of 332 and 56 microM, respectively) whilst the Hairy peptide bound much more strongly (K(i) for dCtBP of 6.22 compared to 133 microM for hCtBP). In addition we have shown that peptides containing identical PXDLS motifs but with different N and C terminal sequences have appreciably different affinities for mammalian CtBP and different structures in solution. We conclude that the factors governing the interactions of CtBPs with partner proteins are more complex than simple possession of the PXDLS motif. In particular the overall secondary structures and amino acid side chains in the binding sites of partner proteins are of importance as well as possible global structural effects in both members of the complex. These data are considered evidence for a multiplicity of CtBPs and partner proteins in the cell.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0006-3002
pubmed:author
pubmed:issnType
Print
pubmed:day
9
pubmed:volume
1546
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
55-70
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11257508-Adenovirus E1A Proteins, pubmed-meshheading:11257508-Alcohol Oxidoreductases, pubmed-meshheading:11257508-Amino Acid Sequence, pubmed-meshheading:11257508-Animals, pubmed-meshheading:11257508-Binding Sites, pubmed-meshheading:11257508-DNA-Binding Proteins, pubmed-meshheading:11257508-Drosophila, pubmed-meshheading:11257508-Enzyme-Linked Immunosorbent Assay, pubmed-meshheading:11257508-Kinetics, pubmed-meshheading:11257508-Magnetic Resonance Spectroscopy, pubmed-meshheading:11257508-Models, Molecular, pubmed-meshheading:11257508-Molecular Sequence Data, pubmed-meshheading:11257508-Peptides, pubmed-meshheading:11257508-Phosphoproteins, pubmed-meshheading:11257508-Repressor Proteins, pubmed-meshheading:11257508-Structure-Activity Relationship, pubmed-meshheading:11257508-Transcription Factors
pubmed:year
2001
pubmed:articleTitle
Structural determinants outside the PXDLS sequence affect the interaction of adenovirus E1A, C-terminal interacting protein and Drosophila repressors with C-terminal binding protein.
pubmed:affiliation
Division of Cancer Studies, University of Birmingham Medical School, Edgbaston, Birmingham B15 2TA, UK. david.molloy@luton.ac.uk
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't