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Investigating Lipase/Stain Interactions: Determining Interfacial Protein Conformation with Surface Spectroscopy.
Saeed, Khezar H; Strunge, Kris; Pedersen, Kasper B; Truelsen, Sigurd F; Christensen, Sune M; Olsen, Lars; Schiøtt, Birgit; Weidner, Tobias.
Afiliación
  • Saeed KH; Department of Chemistry, Aarhus University, Langelandsgade 140, 8000 Aarhus C, Denmark.
  • Strunge K; Department of Chemistry, Aarhus University, Langelandsgade 140, 8000 Aarhus C, Denmark.
  • Pedersen KB; Department of Chemistry, Aarhus University, Langelandsgade 140, 8000 Aarhus C, Denmark.
  • Truelsen SF; Novonesis A/S Biologiens Vej 2, 2800 Kgs Lyngby, Denmark.
  • Christensen SM; Novonesis A/S Biologiens Vej 2, 2800 Kgs Lyngby, Denmark.
  • Olsen L; Novonesis A/S Biologiens Vej 2, 2800 Kgs Lyngby, Denmark.
  • Schiøtt B; Department of Chemistry, Aarhus University, Langelandsgade 140, 8000 Aarhus C, Denmark.
  • Weidner T; Department of Chemistry, Aarhus University, Langelandsgade 140, 8000 Aarhus C, Denmark.
J Phys Chem B ; 128(34): 8162-8169, 2024 Aug 29.
Article en En | MEDLINE | ID: mdl-39158521
ABSTRACT
Conventional bulk protein structure determination methods are not suitable for understanding the distinct and diverse interactions of proteins with interfaces. Notably, interfacial activation is a feature common to many lipases involving movement of a helical "lid" region upon contact with a hydrophobic surface to expose the catalytic site. Here we use the surface specificity of vibrational sum frequency generation spectroscopy (VSFG) spectroscopy to directly probe the conformation of Thermomyces lanuginosus lipase (TLL) at hydrophobic interfaces. The TLL-catalyzed reaction at the air/water interface is monitored by VSFG spectroscopy, showing loss of ester carbonyl modes and appearance of carboxylate stretching modes of the fatty acid products. Furthermore, comparison of experimental and calculated VSFG spectra of the amide I band of TLL allows us to discern the subtle structural changes involved with lid-opening at a hydrophobic surface. Finally, we report a likely orientation of this lid-open state, which interacts with the surface through a loop region away from the lid and active site. This experimental framework for probing protein structure and function at interfaces addresses a significant problem in protein science that is not only impeding the design of better enzymes for biotechnology applications but also drug discovery targeting membrane associated proteins.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Interacciones Hidrofóbicas e Hidrofílicas / Lipasa Idioma: En Revista: J Phys Chem B Asunto de la revista: QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Interacciones Hidrofóbicas e Hidrofílicas / Lipasa Idioma: En Revista: J Phys Chem B Asunto de la revista: QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Estados Unidos