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Titolo:
Elucidation of the binding regions of PAI-1 neutralizing antibodies using chimeric variants of human and rat PAI-1
Autore:
Bijnens, AP; Ngo, TH; Gils, A; Dewaele, J; Knockaert, I; Stassen, JM; Declerck, PJ;
Indirizzi:
Katholieke Univ Leuven, Fac Pharmaceut Sci, Lab Pharmaceut Biol & Phytopharmacol, B-3000 Louvain, Belgium Katholieke Univ Leuven Louvain Belgium B-3000 l, B-3000 Louvain, Belgium Boehringer Ingelheim KG, Cardiovasc Pharmacol, Pharma, Biberach, Germany Boehringer Ingelheim KG Biberach Germany col, Pharma, Biberach, Germany
Titolo Testata:
THROMBOSIS AND HAEMOSTASIS
fascicolo: 5, volume: 85, anno: 2001,
pagine: 866 - 874
SICI:
0340-6245(200105)85:5<866:EOTBRO>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
PLASMINOGEN-ACTIVATOR INHIBITOR-1; MOLECULAR-WEIGHT INHIBITOR; FAST-ACTING INHIBITOR; REACTIVE-CENTER LOOP; HUMAN-PLASMA; TISSUE-TYPE; MONOCLONAL-ANTIBODIES; INTERACTION SITES; IDENTIFICATION; SUBSTRATE;
Keywords:
PAI-1; epitope; chimera; antibody; serpin;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
49
Recensione:
Indirizzi per estratti:
Indirizzo: Declerck, PJ Katholieke Univ Leuven, Fac Pharmaceut Sci, Lab Pharmaceut Biol & Phytopharmacol, Van Evenstr 4, B-3000 Louvain, Belgium Katholieke UnivLeuven Van Evenstr 4 Louvain Belgium B-3000
Citazione:
A.P. Bijnens et al., "Elucidation of the binding regions of PAI-1 neutralizing antibodies using chimeric variants of human and rat PAI-1", THROMB HAEM, 85(5), 2001, pp. 866-874

Abstract

Increased levels of plasminogen activator inhibitor-1 (PAI-1). the main physiological inhibitor of tissue-type plasminogen activator (t-PA) in plasma, are a known risk factor for thromboembolic and cardiovascular diseases. The elucidation of the binding site of inhibitory monoclonal antibodies may contribute to the rational design of PAI-1 modulating therapeutics. In thisstudy. homolog-scanning mutagenesis was used to identify the binding region of a variety of human PAI-I inhibitory antibodies, lacking cross-reactivity with rat PAI-1. Therefore. eight chimeric human/rat PAI-I variants, containing rat PAI-I substitutions at the N-terminal or C-terminal end with splicing sites at positions 26, 81, 187, 277 or 327, were generated and purified. Biochemical characterization revealed that all chimeras were folded properly. Subsequently, surface plasmon resonance was used to determine the affinity of various monoclonal antibodies for these chimera. Comparative analysis of the affinity and ELISA data allowed the identification of the majorbinding region of the inhibitory antibodies MA-8H9D4, MA-33B8F7, MA-44E4. MA-42A2F6 and MA-56A7C10. Thus, three segments in human PAI-1 containing each at least one site involved in the neutralization of PAI-1 activity couldbe identified. i.e. (1) the segment from residue 81 to residue 187 (comprising alpha -helices hD. hE and hF. beta -strands s4C, s3A, s2A and s1A and the loops connecting these elements), (2) the segment between residues 277 and 327 (hl, thIs5A, s5A and s6A) and (3) the region C-terminal from amino acid 327, including the reactive site loop. The current data. together withprevious data, indicate that PAI-I contains at least four different regions that could be considered as putative targets to modulate its activity.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 11/07/20 alle ore 04:13:02