Details

Autor(en) / Beteiligte

• Gonçalves, Karen M.
• Barbosa, Leandro R.S.
• Lima, Luís Maurício T.R.
• Cortines, Juliana R.
• Kalume, Dário E.
• Leal, Ivana C.R.
• Mariz e Miranda, Leandro S.
• de Souza, Rodrigo O.M.
• Cordeiro, Yraima

Titel

Conformational dissection of Thermomyces lanuginosus lipase in solution

Ist Teil von

• Biophysical chemistry, 2014-01, Vol.185, p.88-97

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2014

Quelle

MEDLINE

Beschreibungen/Notizen

• Lipases are triacyl glycerol acyl hydrolases, which catalyze hydrolysis of esters, esterification and transesterification reactions, among others. Some of these enzymes have a large hydrophobic pocket covered by an alpha-helical mobile surface loop (the lid). Protein–protein interactions can occur through adsorption of two open lids of individual lipases. We investigated the conformation and oligomeric state of Thermomyces lanuginosus lipase (TLL) in solution by spectroscopic and mass spectrometry techniques. Information about oligomerization of this important industrial enzyme is only available for TLL crystals; therefore, we have done a throughout investigation of the conformation of this lipase in solution. SDS-PAGE and mass spectrometry analysis of size-exclusion chromatography eluted fractions indicated the presence of both monomeric and dimeric populations of TLL. The stability of the enzyme upon thermal and guanidine hydrochloride treatment was examined by circular dichroism and fluorescence emission spectroscopy. Small angle x-ray scattering and ion mobility mass spectrometry analysis revealed that TLL is found as a mixture of monomers and dimers at the assayed concentrations. Although previous x-ray diffraction data showed TLL as a dimer in the crystal (PDB: 1DT3), to our knowledge our report is the first evidencing that TLL co-exists as stable dimeric and monomeric forms in solution. [Display omitted] •The conformation and oligomeric state of a commercial lipase (TLL) were evaluated.•SAXS data evidenced the presence of monomers and dimers of TLL in solution.•Mass spectrometry analysis confirmed that TLL is present in both forms.•The presence of dimeric species might compromise overall enzyme activity.