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Expression in Escherichia coli, purification, and spectroscopic characterization of two mutant Bet v 1 proteins

Title data

Böhm, Markus ; Rösch, Paul:
Expression in Escherichia coli, purification, and spectroscopic characterization of two mutant Bet v 1 proteins.
In: Biological Chemistry. Vol. 378 (July 1997) Issue 7 . - pp. 687-695.
ISSN 1437-4315
DOI: https://doi.org/10.1515/bchm.1997.378.7.687

Abstract in another language

Bet v 1 is the major birch pollen allergen. A highly efficient expression and purification scheme for mutant forms of this protein was developed on the basis of the pET expression system in order to provide the high quantities of protein needed for spectroscopic and structural work. Bet v 1 (M139L) protein could be purified at high yield (approx. 30 mg from 1 liter of LB medium) in a two-step procedure by the use of metal-affinity chromatography. Matrix assisted laser desorption ionisation mass spectroscopy, and size exclusion chromatography demonstrate the homogeneity and purity of the prepared protein. Spectroscopic methods were used to show that Bet v 1 (M139L) is structurally similar to wild type Bet v 1. Furthermore, we investigated the influence of the nature of amino acid 139 on the thermodynamic behaviour of the protein by replacing the leucine residue by alanine. While there appears to be no global structural effect of this mutation, the thermostability of Bet v 1 is greatly decreased.

Further data

Item Type: Article in a journal
Refereed: Yes
Institutions of the University: Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Former Professors > Chair Biopolymers - Univ.-Prof. Dr. Paul Rösch
Faculties
Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Former Professors
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biopolymers
Result of work at the UBT: Yes
DDC Subjects: 500 Science > 540 Chemistry
500 Science > 570 Life sciences, biology
Date Deposited: 15 Jan 2019 12:36
Last Modified: 16 May 2019 05:37
URI: https://eref.uni-bayreuth.de/id/eprint/46895