Bioincorporation of telluromethionine into proteins: a promising new approach for X-ray structure analysis of proteins
| Autor: | N, Budisa, W, Karnbrock, S, Steinbacher, A, Humm, L, Prade, T, Neuefeind, L, Moroder, R, Huber |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Amidinotransferases
Glycoside Hydrolases Protein Conformation Circular Dichroism DNA-Directed RNA Polymerases Viral Tail Proteins Crystallography X-Ray Recombinant Proteins Viral Proteins Matrix Metalloproteinase 8 Methionine Fermentation Humans Collagenases Annexin A5 Tellurium Glutathione Transferase |
| Zdroj: | Journal of molecular biology. 270(4) |
| ISSN: | 0022-2836 |
| Popis: | A simple and efficient method for the specific and quantitative replacement of the naturally occurring amino acid methionine by its isosteric analogue telluromethionine in the expression of recombinant proteins has been developed. The method requires a controlable and competitive expression system like the bacteriophage T7 polymerase/promoter in a methionine-auxotrophic host. Using methionine-auxotrophic Escherichia coli strains, incorporation of telluromethionine at high yields has been achieved for human recombinant annexin V, human mitochondrial transamidase, Arabidopsis glutathione-S-transferase and the N-terminal domain of Salmonella tailspike adhesion protein as confirmed by amino acid, mass-spectrometric and X-ray analyses. Expressed and purified telluromethionine-proteins and native proteins were found to crystallise isomorphously. In terms of efficient bio-expression, isomorphism of crystals and relative abundance of methionine residues, the production of telluromethionine-proteins as heavy-atom derivatives offers a valid and general approach in X-ray analysis by the method of multiple isomorphous replacement. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect