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Pregled bibliografske jedinice broj: 263566

Mass spectrometric evidence of covalently-bound tetrahydrolipstatin at the catalytic serine of Streptomyces rimosus lipase


Leščić Ašler, Ivana; Zehl, Martin; Kovačić, Filip; Müller, Roland; Abramić, Marija; Allmaier, Günter; Kojić-Prodić, Biserka
Mass spectrometric evidence of covalently-bound tetrahydrolipstatin at the catalytic serine of Streptomyces rimosus lipase // Biochimica et Biophysica Acta (BBA) - General Subjects, 1770 (2007), 2; 163-170 (međunarodna recenzija, članak, znanstveni)


Naslov
Mass spectrometric evidence of covalently-bound tetrahydrolipstatin at the catalytic serine of Streptomyces rimosus lipase

Autori
Leščić Ašler, Ivana ; Zehl, Martin ; Kovačić, Filip ; Müller, Roland ; Abramić, Marija ; Allmaier, Günter ; Kojić-Prodić, Biserka

Izvornik
Biochimica et Biophysica Acta (BBA) - General Subjects (0304-4165) 1770 (2007), 2; 163-170

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

Ključne riječi
Streptomyces rimosus; extracellular SGNH-lipase; tetrahydrolipstatin; covalent inhibition; MALDI tandem mass spectrometry; ESI mass spectrometry; capillary-gel-electrophoresis-on-the-chip; kinetics

Sažetak
Recently, we detected that the lipase from Streptomyces rimosus belongs to a huge but poorly characterised family of SGNH hydrolases having the alpha/beta/alpha-fold. Our biochemical characterisation is related to the specific inhibition of an extracellular lipase from Streptomyces rimosus (SRL, 24.2 kDa, Q93MW7) by preincubation method with tetrahydrolipstatin (THL). In high molar excess, THL/SRL = 590 at 25 &ordm ; ; ; C, pH = 7.0, after 2 h of incubation in an aqueous system, 56 % of the enzyme inhibition was reached, whereas at the same conditions in the presence of 50 % (v/v) 2-propanol/water 71 % enzyme inhibition was obtained. Kinetics measurements are in agreement with pseudo-first-order kinetics. The nucleophilic attack of the catalytic serine 10 of SRL results by an opening of beta-lactam ring of tetrahydrolipstatin and formation of a covalent ester bond. The intact covalent complex of SRL-inhibitor was analysed by ESI and vacuum MALDI mass spectrometry and furthermore the exact covalent THL linkage was determined by vacuum MALDI high energy collision induced dissociation tandem mass spectrometric experiments.

Izvorni jezik
Engleski

Znanstvena područja
Kemija



POVEZANOST RADA


Projekt / tema
098-1191344-2938 - Molekularna enzimologija i proteinske interakcije hidrolaza (Marija Abramić, )
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


Uključenost u ostale bibliografske baze podataka:


  • Chemical Abstracts