Document Detail


Powerful binders for the D-dimer by conjugation of the GPRP peptide to polypeptides from a designed set - illustrating a general route to new binders for proteins.
MedLine Citation:
PMID:  23151063     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
The synthetic tetrapeptide GPRP based on the amino-terminal GPR sequence of the fibrin α-chain, binds the D-dimer protein with a dissociation constant KD of 25 μM. The D-dimer protein, a well-known biomarker for thrombosis, contains two cross-linked D fragments from the fibrinogen protein formed upon degradation of the fibrin gel, the core component of blood clots. In order to develop a specific high-affinity binder for the D-dimer protein, GPRP was conjugated via an aliphatic spacer to each member of a set of sixteen polypeptides designed for the development of binder molecules for proteins in general. The binders were individually characterised and ranked using surface plasmon resonance (SPR) analysis. The dissociation constant of the complex formed from the D-dimer and 4-D15L8-GPRP labelled with fluorescein was determined by fluorescense titration and found to be 3 nM, an affinity four orders of magnitude higher than that of free GPRP. According to SPR analysis binding was completely inhibited by free GPRP at mM concentrations and the polypeptide conjugate was therefore shown to bind specifically to the binding site of GPRP. Affinities were further enhanced by dimerisation of the polypeptide conjugates via a bifunctional linker resulting in dissociation constants that were further decreased (affinities increased) by factors of 2-4. The results suggest an efficient route to specific binders for proteins based on short peptides with affinites that need only to be modest, thus shortening the time of binder development dramatically.
Authors:
Ramesh Ramapanicker; Xiaojiao Sun; Johan Viljanen; Lars Baltzer
Related Documents :
22238323 - Aβ delays fibrin clot lysis by altering fibrin structure and attenuating plasminogen b...
6206963 - Effects of bay k 8644, a dihydropyridine analog, on [3h]nitrendipine binding to canine ...
24032403 - Differential regulation of camkiiα interactions with mglur5 and nmda receptors by ca(2...
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-11-14
Journal Detail:
Title:  Bioconjugate chemistry     Volume:  -     ISSN:  1520-4812     ISO Abbreviation:  Bioconjug. Chem.     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-15     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9010319     Medline TA:  Bioconjug Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  A Solid State Density Functional Study of Crystalline Thiophene-Based Oligomers and Polymers.
Next Document:  Glycoluril dimer isomerization under aqueous acidic conditions related to cucurbituril formation.