Transition of ovalbumin to thermostable structure entails conformational changes involving the reactive center loop
| Autor: | Ikunosin Kato, Hiroshi Shinohara, Mamoru Sato, Masahisa Horiuchi, Yasushi Sugimoto, Yuji Minami, Takayoshi Aoki, Katsumi Koga, Junichi Kurisaki, Takahiro Kusakabe |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Proteases
Calorimetry Differential Scanning biology Ovalbumin Protein Conformation Chemistry Stereochemistry Hydrolysis Biophysics Subtilisin respiratory system Serpin Biochemistry Epitope Dephosphorylation Residue (chemistry) Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Enzyme Stability biology.protein Phosphorylation Molecular Biology Reactive center |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - General Subjects. 1770:5-11 |
| ISSN: | 0304-4165 |
| DOI: | 10.1016/j.bbagen.2006.06.019 |
| Popis: | Ovalbumin is a serpin without inhibitory activity against proteases. During embryonic development, ovalbumin in the native (N) form undergoes changes and takes a heat-stable form, which was previously named HS-ovalbumin. It has been known that N-ovalbumin is artificially converted to another thermostable form called S-ovalbumin by heating at an alkaline pH. Here, we characterized further the three ovalbumin forms, N, HS, and S. The epitope of the monoclonal antibody 2B3/2H11, which recognizes N- and HS-ovalbumin but not S-ovalbumin, was found to reside in the region Glu-Val-Val-Gly-Ala-Ser-Glu-Ala-Gly-Val-Asp-Ala-Ala-Ser-Val-Ser-Glu-Glu-Phe-Arg, which corresponds to 340-359 of amino acid residues and is contained in the reactive center loop (RCL). Removal of RCL by elastase or subtilisin mitigated binding of the antibody. Dephosphorylation experiments indicated that the phosphorylated Ser-344 residue located on RCL is crucial for the epitope recognition. We suggest that the shift to the heat-stable form of ovalbumin accompanies a movement of RCL. |
| Databáze: | OpenAIRE |
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