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Inhibition of extracellular lipase from Streptomyces rimosus with 3, 4-dichloroisocoumarin


Leščić Ašler, Ivana; Kovačić, Filip; Marchetti-Deschmann, Martina; Allmaier, Günter; Štefanić, Zoran; Kojić-Prodić, Biserka
Inhibition of extracellular lipase from Streptomyces rimosus with 3, 4-dichloroisocoumarin // Journal of enzyme inhibition and medicinal chemistry, 28 (2013), 5; 1094-1104 doi:10.3109/14756366.2012.716834 (međunarodna recenzija, članak, znanstveni)


Naslov
Inhibition of extracellular lipase from Streptomyces rimosus with 3, 4-dichloroisocoumarin

Autori
Leščić Ašler, Ivana ; Kovačić, Filip ; Marchetti-Deschmann, Martina ; Allmaier, Günter ; Štefanić, Zoran ; Kojić-Prodić, Biserka

Izvornik
Journal of enzyme inhibition and medicinal chemistry (1475-6366) 28 (2013), 5; 1094-1104

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

Ključne riječi
Enzyme inhibition kinetics; active-site titration; mass spectrometry; SGNH-hydrolase

Sažetak
Kinetic characterization of lipase inhibition was performed by activity measurement and mass spectrometry (MS), for the first time with serine protease inhibitor 3, 4-dichloroisocoumarin (DCI). Inhibition of Streptomyces rimosus extracellular lipase (SrLip), a member of the SGNH superfamily, by means of DCI follows the mechanism of two-step irreversible inhibition. The dissociation constant of the noncovalent EI complex and first-order rate constant for inactivation were determined by incubation (Ki* = 26.6 ± 2.8 μM, k2 = 12.2 ± 0.6 min–1) or progress curve (Ki* = 6.5 ± 1.5 μM, k2 = 0.11 ± 0.01 min–1) method. Half-times of reactivation for lipase inhibited with 10-fold molar excess of DCI were determined by activity measurement (t1/2 = 11.3 ± 0.2 h), matrix-assisted laser desorption/ionization (MALDI, t1/2 = 13.5 ± 0.4 h), and electro-spray ionization (ESI, t1/2 = 12.2 ± 0.5 h) MS. The active SrLip concentration was determined by incubating the enzyme with near equimolar concentrations of DCI, followed by activity and MS measurement.

Izvorni jezik
Engleski

Znanstvena područja
Kemija



POVEZANOST RADA


Projekt / tema
098-1191344-2943 - Protein-ligand međudjelovanja na atomnoj razini (Marija Luić, )

Ustanove
Institut "Ruđer Bošković", Zagreb

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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