| Autor: |
Eidsness MK; Department of Chemistry, University of Georgia, Athens 30602., O'Dell SE, Kurtz DM Jr, Robson RL, Scott RA |
| Jazyk: |
angličtina |
| Zdroj: |
Protein engineering [Protein Eng] 1992 Jun; Vol. 5 (4), pp. 367-71. |
| DOI: |
10.1093/protein/5.4.367 |
| Abstrakt: |
A synthetic gene based on the published amino acid sequence for Clostridium pasteurianum rubredoxin was constructed, cloned in Escherichia coli 71/18 and expressed using the T7 RNA polymerase/promoter system in E. coli HMS273. UV/visible spectroscopy and metal analyses indicated that the as-isolated synthetic gene product is a mixture of holo-(i.e. iron-containing) rubredoxin and zinc-substituted rubredoxin, with the latter amounting to approximately 70% of the total rubredoxin. The UV/visible absorption and resonance Raman spectra of the cloned holorubredoxin are characteristic of the native rubredoxin-type iron site. N-terminal amino acid sequencing suggests that the gene product consists of at least three polypeptide species with the initial sequences (approximate relative abundances): Met-Met-Lys-... (63%), blocked (30%) and Met-Lys-... (7%). The blocked portion presumably consists of a mixture of nMet-Met-Lys-... and nMet-Lys-..., where nMet represents an amino-blocked methionine residue. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
