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Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents


Karačić, Zrinka; Špoljarić, Jasminka; Rožman, Marko; Abramić, Marija
Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents // Biological chemistry, 393 (2012), 12; 1523-1532 doi:10.1515/hsz-2012-0181 (međunarodna recenzija, članak, znanstveni)


Naslov
Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents

Autori
Karačić, Zrinka ; Špoljarić, Jasminka ; Rožman, Marko ; Abramić, Marija

Izvornik
Biological chemistry (1431-6730) 393 (2012), 12; 1523-1532

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Glutathionylation; mass spectrometry; metallopeptidase M49; reactive cysteines; sulfhydryl reagent

Sažetak
Human dipeptidyl peptidase III (DPP III) is a member of the metallopeptidase family M49, involved in protein metabolism and oxidative stress response. DPP III crystal structure shows the two lobe-like domains separated by a wide cleft. The human enzyme has a total of six cysteines, three in the lower (Cys19, Cys147, and Cys176) and three in the upper (Cys509, Cys519, and Cys654), catalytic, domain containing the active site zinc ion. To elucidate the molecular basis of this enzyme’s susceptibility to sulfhydryl reagents, biochemical analysis of a set of Cys to Ala mutants was used, supported by mass spectrometry. Cys176, a residue 44 Å apart from the catalytic center of the ligand-free enzyme, was found responsible for the inactivation with the submicromolar concentration of an organomercurial compound, and three additional cysteines contributed to sensitivity to aromatic disulfides. Upon treatment with oxidized glutathione [glutathione disulfide (GSSG)], cysteine residues at positions 147, 176, and 654 were found glutathionylated. The mutational analysis confirmed the involvement of Cys176 and Cys654 in human DPP III inactivation by GSSG. Observation that Cys176, a residue quite distant from the active center, contributes to enzyme inactivation, indicates that the substrate-binding site of human DPP III comprises both lower and upper protein domain.

Izvorni jezik
Engleski

Znanstvena područja
Kemija



POVEZANOST RADA


Projekt / tema
098-0982915-2945 - Spektroskopija, kemijska svojstva i reakcije biološki aktivnih molekula (Branka Kovač, )
098-1191344-2938 - Molekularna enzimologija i proteinske interakcije hidrolaza (Marija Abramić, )

Ustanove
Institut "Ruđer Bošković", Zagreb

Citiraj ovu publikaciju

Karačić, Zrinka; Špoljarić, Jasminka; Rožman, Marko; Abramić, Marija
Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents // Biological chemistry, 393 (2012), 12; 1523-1532 doi:10.1515/hsz-2012-0181 (međunarodna recenzija, članak, znanstveni)
Karačić, Z., Špoljarić, J., Rožman, M. & Abramić, M. (2012) Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents. Biological chemistry, 393 (12), 1523-1532 doi:10.1515/hsz-2012-0181.
@article{article, year = {2012}, pages = {1523-1532}, DOI = {10.1515/hsz-2012-0181}, keywords = {glutathionylation, mass spectrometry, metallopeptidase M49, reactive cysteines, sulfhydryl reagent}, journal = {Biological chemistry}, doi = {10.1515/hsz-2012-0181}, volume = {393}, number = {12}, issn = {1431-6730}, title = {Molecular determinants of human dipeptidyl peptidase III sensitivity to thiol modifying reagents}, keyword = {glutathionylation, mass spectrometry, metallopeptidase M49, reactive cysteines, sulfhydryl reagent} }

Časopis indeksira:


  • Current Contents Connect (CCC)
  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus
  • MEDLINE


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