The Interaction of Fusarium solani pisi Cutinase with Long Chain Spin Label Esters
| Autor: | Kristian Raaby Poulsen, Thomas Kjær Sørensen, Laurent Duroux, Steffen B. Petersen, Evamaria I. Petersen, Reinhard Wimmer |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Cutinase
Protein Folding Free Radicals Stereochemistry Biochemistry Protein Structure Secondary Substrate Specificity Cyclic N-Oxides Fungal Proteins Fusarium Serine Side chain Spin label Nuclear Magnetic Resonance Biomolecular Micelles chemistry.chemical_classification Alanine biology Electron Spin Resonance Spectroscopy Substrate (chemistry) Active site Esters biology.organism_classification Amino acid chemistry biology.protein Spin Labels Oxyanion hole Carboxylic Ester Hydrolases Fusarium solani |
| Zdroj: | Aalborg University Poulsen, K R, Sørensen, T K, Duroux, L, Petersen, E, Petersen, S B & Wimmer, R 2006, ' The interaction of Fusarium solani pisi cutinase with long chain spin label esters ', Biochemistry, vol. 45, no. 30, pp. 9163-9171 . |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi060329i |
| Popis: | We here present a study of the interaction between the Fusarium solani pisi cutinase mutant S120A and spin-labeled 4,4-dimethyloxazoline-N-oxyl-(DOXYL)-stearoyl-glycerol substrates in a micellar system. The interaction is detected by NMR measuring changes in chemical shift for 1H and 15N as well as relaxation parameters for backbone 1H (T1) and 15N (T1, T2) atoms as well as for side chain methyl groups 1H (T1). The detected interaction shows a weak binding of cutinase to the lipid micelles. Structural and mobility changes are located inside and around the active site, its flanking loops, and the oxyanion hole, respectively. Relaxation changes in the amino acid pairs Ser 92, Ala 93 and Thr 173, Gly 174 positioned at the edge of each of the active site flanking loops make these residues prime candidates for hinges, allowing for structural rearrangement during substrate binding. The cutinase mutant S120A used carries a 15 amino acid pro-peptide; the significance of this pro-peptide was so far undetermined. We show here that the pro-peptide is affected by the presence of the micellar substrate. Relaxation enhancements indicative of spatial proximity between the DOXYL group in the lipid chain and some hydrophobic residues surrounding the active site could be found. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|