Počet záznamů: 1

Structural and biochemical characterization of the folyl-poly-gamma-L-glutamate hydrolyzing activity of human glutamate carboxypeptidase II

SYSNO ASEP	0431487
Druh ASEP	J - Článek v odborném periodiku
Zařazení RIV	J - Článek v odborném periodiku
Poddruh J	Článek ve WOS
Název	Structural and biochemical characterization of the folyl-poly-gamma-L-glutamate hydrolyzing activity of human glutamate carboxypeptidase II
Tvůrce(i)	Navrátil, Michal (UOCHB-X)_RID Ptáček, Jakub (BTO-N)_RID Šácha, Pavel (UOCHB-X)_{RID, ORCID} Starková, Jana (UOCHB-X) Lubkowski, J. (US) Bařinka, Cyril (BTO-N)_{RID, ORCID} Konvalinka, Jan (UOCHB-X)_{RID, ORCID}
Celkový počet autorů	7
Zdroj.dok.	FEBS Journal - ISSN 1742-464X Roč. 281, č. 14 (2014), s. 3228-3242
Poč.str.	15 s.
Jazyk dok.	eng - angličtina
Země vyd.	GB - Velká Británie
Klíč. slova	arene-binding site ; crystal structure ; folate hydrolase 1 ; H475Y(1561C -> T) ; polymorphism ; zinc metalloprotease
Vědní obor RIV	CE - Biochemie
CEP	GAP304/12/0847 GA ČR - Grantová agentura ČR
	ED1.1.00/02.0109 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy
Institucionální podpora	UOCHB-X - RVO:61388963 ; BTO-N - RVO:86652036
UT WOS	000339800800011
EID SCOPUS	84904732507
DOI	10.1111/febs.12857
Anotace	In addition to its well-characterized role in the central nervous system, human glutamate carboxypeptidase II (GCPII; Uniprot ID Q04609) acts as a folate hydrolase in the small intestine, participating in the absorption of dietary polyglutamylated folates (folyl-n-gamma-L-glutamic acid), which are the provitamin form of folic acid (also known as vitamin B-9). Despite the role of GCPII as a folate hydrolase, nothing is known about the processing of polyglutamylated folates by GCPII at the structural or enzymological level. Moreover, many epidemiologic studies on the relationship of the naturally occurring His475Tyr polymorphism to folic acid status suggest that this polymorphism may be associated with several pathologies linked to impaired folate metabolism. In the present study, we report: (a) a series X-ray structures of complexes between a catalytically inactive GCPII mutant (Glu424Ala) and a panel of naturally occurring polyglutamylated folates; (b) the X-ray structure of the His475Tyr variant at a resolution of 1.83 angstrom; (c) the study of the recently identified arene-binding site of GCPII through mutagenesis (Arg463Leu, Arg511Leu and Trp541Ala), inhibitor binding and enzyme kinetics with polyglutamylated folates as substrates; and (d) a comparison of the thermal stabilities and folate-hydrolyzing activities of GCPII wild-type and His475Tyr variants. As a result, the crystallographic data reveal considerable details about the binding mode of polyglutamylated folates to GCPII, especially the engagement of the arene binding site in recognizing the folic acid moiety. Additionally, the combined structural and kinetic data suggest that GCPII wild-type and His475Tyr variant are functionally identical.
Pracoviště	Ústav organické chemie a biochemie
Kontakt	asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434
Rok sběru	2015

Počet záznamů: 1