Structural, functional and chemical changes in Pseudozyma antarctica lipase B on exposure to hydrogen peroxide

Biochimie
Ulrika TörnvallRajni Hatti-Kaul

Abstract

The effect on primary, secondary, tertiary and quaternary structure of Pseudozyma (formerly Candida) antarctica lipase B (PalB) on exposure to hydrogen peroxide was investigated using nano-electrospray ionization-mass spectrometry (nano-ESI-MS), liquid chromatography tandem mass spectrometry (LC/MS/MS), circular dichroism (CD), and dynamic light scattering (DLS). Treatment with hydrogen peroxide generated heavier protein variants, with a mass gain that increased with increasing incubation time. Furthermore, elevated concentration of H(2)O(2) was shown to result in partial fragmentation of the protein. Proteolytic digestion of the enzyme gave primary sequence coverage of more than 90%, revealing oxidation of methionine, tryptophan and cystine residues. The active site histidine was not observed in oxidized form in any of the experiments. However, oxidation of cystine to cysteic acid indicated disruption of disulphide bridges, and CD evaluations confirmed that severe changes to the secondary structure towards random coil had occurred. The structural changes could be an effect of the observed amino acid side chain oxidations, and was correlated with deactivation of the lipase. From DLS experiments, it was seen that the lipase expo...Continue Reading

References

Jan 1, 1995·Free Radical Biology & Medicine·W Vogt
Jan 1, 1996·Rapid Communications in Mass Spectrometry : RCM·F M LagerwerfJ Haverkamp
Aug 15, 1997·The Journal of Biological Chemistry·B S Berlett, E R Stadtman
Mar 25, 1998·Biophysical Journal·J GaoT C Squier
May 18, 2000·Accounts of Chemical Research·K L Schey, E L Finley
Apr 3, 2001·Protein Expression and Purification·J C Rotticci-MulderM Martinelle
Oct 26, 2001·The Journal of Biological Chemistry·R GrimaudF Barras
Jun 12, 2004·The Journal of Dairy Research·Lars Wiking, Jacob H Nielsen
Aug 6, 2004·The Journal of Biological Chemistry·Thea van der WijkJeroen den Hertog
Feb 1, 2005·Biochimica Et Biophysica Acta·Michael J Davies
May 11, 2005·Chembiochem : a European Journal of Chemical Biology·Anders O MagnussonKarl Hult
Jun 30, 2006·Biotechnology and Bioengineering·Inga PetryPeter J Halling
Aug 7, 2007·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·David LöfAlex Evilevitch
Nov 7, 2007·Rapid Communications in Mass Spectrometry : RCM·Sanjay VenkateshJoshua S Sharp
May 15, 2008·Applied Biochemistry and Biotechnology·Abhijeet P Borole, Brian H Davison
Jul 4, 2009·Rapid Communications in Mass Spectrometry : RCM·Sofia GuedesPedro Domingues
Aug 15, 2009·Rapid Communications in Mass Spectrometry : RCM·Ulrika TörnvallMartin Hedström
Feb 4, 2010·Journal of Proteome Research·Ashraf G Madian, Fred E Regnier
Feb 6, 2010·Proceedings of the National Academy of Sciences of the United States of America·Yuan Qi WongMichael D W Griffin
Mar 27, 2010·Analytical Chemistry·Brian C GauMichael L Gross

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