• Je něco špatně v tomto záznamu ?

Prolyl Oligopeptidase from the Blood Fluke Schistosoma mansoni: From Functional Analysis to Anti-schistosomal Inhibitors

P. Fajtová, S. Štefanić, M. Hradilek, J. Dvořák, J. Vondrášek, A. Jílková, L. Ulrychová, JH. McKerrow, CR. Caffrey, M. Mareš, M. Horn,

. 2015 ; 9 (6) : e0003827. [pub] 20150603

Jazyk angličtina Země Spojené státy americké

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc16010133

BACKGROUND: Blood flukes of the genus Schistosoma cause schistosomiasis, a parasitic disease that infects over 240 million people worldwide, and for which there is a need to identify new targets for chemotherapeutic interventions. Our research is focused on Schistosoma mansoni prolyl oligopeptidase (SmPOP) from the serine peptidase family S9, which has not been investigated in detail in trematodes. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrate that SmPOP is expressed in adult worms and schistosomula in an enzymatically active form. By immunofluorescence microscopy, SmPOP is localized in the tegument and parenchyma of both developmental stages. Recombinant SmPOP was produced in Escherichia coli and its active site specificity investigated using synthetic substrate and inhibitor libraries, and by homology modeling. SmPOP is a true oligopeptidase that hydrolyzes peptide (but not protein) substrates with a strict specificity for Pro at P1. The inhibition profile is analogous to those for mammalian POPs. Both the recombinant enzyme and live worms cleave host vasoregulatory, proline-containing hormones such as angiotensin I and bradykinin. Finally, we designed nanomolar inhibitors of SmPOP that induce deleterious phenotypes in cultured schistosomes. CONCLUSIONS/SIGNIFICANCE: We provide the first localization and functional analysis of SmPOP together with chemical tools for measuring its activity. We briefly discuss the notion that SmPOP, operating at the host-parasite interface to cleave host bioactive peptides, may contribute to the survival of the parasite. If substantiated, SmPOP could be a new target for the development of anti-schistosomal drugs.

Citace poskytuje Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc16010133
003      
CZ-PrNML
005      
20160418102207.0
007      
ta
008      
160408s2015 xxu f 000 0|eng||
009      
AR
024    7_
$a 10.1371/journal.pntd.0003827 $2 doi
024    7_
$a 10.1371/journal.pntd.0003827 $2 doi
035    __
$a (PubMed)26039195
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a xxu
100    1_
$a Fajtová, Pavla $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic; First Faculty of Medicine, Charles University, Prague, Czech Republic.
245    10
$a Prolyl Oligopeptidase from the Blood Fluke Schistosoma mansoni: From Functional Analysis to Anti-schistosomal Inhibitors / $c P. Fajtová, S. Štefanić, M. Hradilek, J. Dvořák, J. Vondrášek, A. Jílková, L. Ulrychová, JH. McKerrow, CR. Caffrey, M. Mareš, M. Horn,
520    9_
$a BACKGROUND: Blood flukes of the genus Schistosoma cause schistosomiasis, a parasitic disease that infects over 240 million people worldwide, and for which there is a need to identify new targets for chemotherapeutic interventions. Our research is focused on Schistosoma mansoni prolyl oligopeptidase (SmPOP) from the serine peptidase family S9, which has not been investigated in detail in trematodes. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrate that SmPOP is expressed in adult worms and schistosomula in an enzymatically active form. By immunofluorescence microscopy, SmPOP is localized in the tegument and parenchyma of both developmental stages. Recombinant SmPOP was produced in Escherichia coli and its active site specificity investigated using synthetic substrate and inhibitor libraries, and by homology modeling. SmPOP is a true oligopeptidase that hydrolyzes peptide (but not protein) substrates with a strict specificity for Pro at P1. The inhibition profile is analogous to those for mammalian POPs. Both the recombinant enzyme and live worms cleave host vasoregulatory, proline-containing hormones such as angiotensin I and bradykinin. Finally, we designed nanomolar inhibitors of SmPOP that induce deleterious phenotypes in cultured schistosomes. CONCLUSIONS/SIGNIFICANCE: We provide the first localization and functional analysis of SmPOP together with chemical tools for measuring its activity. We briefly discuss the notion that SmPOP, operating at the host-parasite interface to cleave host bioactive peptides, may contribute to the survival of the parasite. If substantiated, SmPOP could be a new target for the development of anti-schistosomal drugs.
650    _2
$a zvířata $7 D000818
650    _2
$a katalytická doména $x genetika $7 D020134
650    _2
$a DNA primery $x genetika $7 D017931
650    _2
$a Escherichia coli $7 D004926
650    _2
$a stanovení celkové genové exprese $7 D020869
650    _2
$a regulace genové exprese enzymů $x fyziologie $7 D015971
650    _2
$a hydrolýza $7 D006868
650    _2
$a imunoblotting $7 D015151
650    _2
$a fluorescenční mikroskopie $7 D008856
650    12
$a molekulární modely $7 D008958
650    _2
$a kvantitativní polymerázová řetězová reakce $7 D060888
650    _2
$a rekombinantní proteiny $x genetika $x metabolismus $7 D011994
650    _2
$a Schistosoma mansoni $x enzymologie $7 D012550
650    _2
$a serinové endopeptidasy $x genetika $x metabolismus $7 D012697
650    _2
$a substrátová specifita $7 D013379
655    _2
$a časopisecké články $7 D016428
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Štefanić, Saša $u Institute of Parasitology, University of Zurich, Zurich, Switzerland.
700    1_
$a Hradilek, Martin $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
700    1_
$a Dvořák, Jan $u Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic; Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech Republic, Ceske Budejovice, Czech Republic.
700    1_
$a Vondrášek, Jiří $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
700    1_
$a Jílková, Adéla $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
700    1_
$a Ulrychová, Lenka $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic; Faculty of Science, Charles University, Prague, Czech Republic.
700    1_
$a McKerrow, James H $u Center for Innovation and Discovery in Parasitic Diseases, Department of Pathology, University of California, San Francisco, San Francisco, California, United States of America.
700    1_
$a Caffrey, Conor R $u Center for Innovation and Discovery in Parasitic Diseases, Department of Pathology, University of California, San Francisco, San Francisco, California, United States of America.
700    1_
$a Mareš, Michael $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
700    1_
$a Horn, Martin $u Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
773    0_
$w MED00165375 $t PLoS neglected tropical diseases $x 1935-2735 $g Roč. 9, č. 6 (2015), s. e0003827
856    41
$u https://pubmed.ncbi.nlm.nih.gov/26039195 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20160408 $b ABA008
991    __
$a 20160418102254 $b ABA008
999    __
$a ok $b bmc $g 1113562 $s 934501
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2015 $b 9 $c 6 $d e0003827 $e 20150603 $i 1935-2735 $m PLoS neglected tropical diseases $n PLoS negl. trop. dis. $x MED00165375
LZP    __
$a Pubmed-20160408

Najít záznam

Citační ukazatele

Možnosti archivace