1GT6

S146A mutant of Thermomyces (Humicola) lanuginosa lipase complex with oleic acid

Summary for 1GT6

• Entry DOI: 10.2210/pdb1gt6/pdb
• Related: 1DT3 1DT5 1DTE 1DU4
• Descriptor: Lipase, OLEIC ACID (3 entities in total)
• Functional Keywords: lipase, hydrolase, lipid degradation, zymogen
• Biological source: Thermomyces lanuginosus (Humicola lanuginosa)
• Total number of polymer chains: 2
• Total formula weight: 59217.89

Authors

Brzozowski, A.M.,Yapoudjian, S.,Ivanova, M.G.,Patkar, S.A.,Vind, J.,Svendsen, A.,Verger, R. (deposition date: 2002-01-11, release date: 2003-01-23, Last modification date: 2024-10-16)

Primary citation

Yapoudjian, S.,Ivanova, M.G.,Brzozowski, A.M.,Patkar, S.A.,Vind, J.,Svendsen, A.,Verger, R.
Binding of Thermomyces (Humicola) lanuginosa lipase to the mixed micelles of cis-parinaric acid/NaTDC.
Eur.J.Biochem., 269:1613-1621, 2002

PubMed Abstract: The binding of Thermomyces lanuginosa lipase and its mutants [TLL(S146A), TLL(W89L), TLL(W117F, W221H, W260H)] to the mixed micelles of cis-parinaric acid/sodium taurodeoxycholate at pH 5.0 led to the quenching of the intrinsic tryptophan fluorescence emission (300-380 nm) and to a simultaneous increase in the cis-parinaric acid fluorescence emission (380-500 nm). These findings were used to characterize the Thermomyces lanuginosa lipase/cis-parinaric acid interactions occurring in the presence of sodium taurodeoxycholate. The fluorescence resonance energy transfer and Stern-Volmer quenching constant values obtained were correlated with the accessibility of the tryptophan residues to the cis-parinaric acid and with the lid opening ability of Thermomyces lanuginosa lipase (and its mutants). TLL(S146A) was found to have the highest fluorescence resonance energy transfer. In addition, a TLL(S146A)/oleic acid complex was crystallised and its three-dimensional structure was solved. Surprisingly, two possible binding modes (sn-1 and antisn1) were found to exist between oleic acid and the catalytic cleft of the open conformation of TLL(S146A). Both binding modes involved an interaction with tryptophan 89 of the lipase lid, in agreement with fluorescence resonance energy transfer experiments. As a consequence, we concluded that TLL(S146A) mutant is not an appropriate substitute for the wild-type Thermomyces lanuginosa lipase for mimicking the interaction between the wild-type enzyme and lipids.

PubMed: 11895431
DOI: 10.1046/j.1432-1327.2002.02786.x

Experimental method

X-RAY DIFFRACTION (2.2 Å)