Thermomyces lanuginosus lipase in the liquid-crystalline phases of aqueous phytantriol: X-ray diffraction and vibrational spectroscopic studies.
The influence of Thermomyces lanuginosus lipase (TLL) on the phase behaviour of liquid-crystalline phases of aqueous phytantriol as well as conformational changes of TLL entrapped in the cubic Q230 phase have been studied by small angle X-ray diffraction (SAXD), FT-Raman, and FT-IR techniques. It was found that the lipidic Q230 phase is able to accommodate up to 10 wt.% of TLL, and the temperature of phase transition to the inverted hexagonal phase H(II) increases indicating stabilizing effect of the protein. FT-Raman analysis of Trp amino acid marker band W3 revealed that the average rotation angle around the C3-Cbeta bond of four Trp residues of TLL in the Q230 phase increases. Reasoning from available TLL crystallographic data, this result is explained by structural transition of entrapped protein to so-called "open" and more related to the enzymatically-active conformation. TLL secondary structure analysis by amide I and amide III vibrational bands showed that content of alpha-helixes does not change, while a part of beta-sheet structures transforms to less ordered elements upon incorporation of protein into the Q230 phase of aqueous phytantriol.
Misiūnas
Audrius
A
Institute of Biochemistry, Mokslininku 12, LT-08662 Vilnius, Lithuania. audriusm@bchi.lt
Talaikyte
Zita
Z
Niaura
Gediminas
G
Razumas
Valdemaras
V
Nylander
Tommy
T
D000577:Amides / Q000737:chemistry; D001203:Ascomycota / Q000201:enzymology*; D003460:Crystallization; D004220:Disulfides / Q000737:chemistry; D005233:Fatty Alcohols / Q000737:chemistry*; D008049:Lipase / Q000378:metabolism*; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D008958:Models, Molecular; D015394:Molecular Structure; D044367:Phase Transition; D013055:Spectrophotometry, Infrared; D013059:Spectrum Analysis, Raman; D013696:Temperature; D014364:Tryptophan / Q000737:chemistry; D014732:Vibration; D014867:Water / Q000737:chemistry; D014961:X-Ray Diffraction
D000577:Amides; D004220:Disulfides; D005233:Fatty Alcohols; D008055:Lipids; D014867:Water; D014364:Tryptophan; C508873:3,7,11,15-tetramethyl-1,2,3-hexadecanetriol; D008049:Lipase