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Identification of the Amino Acid Residue Modified in Bacillus stearothermophilus Alcohol Dehydrogenase by the NAD+ Analogue 4-(3-Bromoacetylpyridinio)butyldiphosphoadenosine
| Content Provider | Scilit |
|---|---|
| Author | Jeck, Reinhard Woenckhaus, Christoph Harris, J. Ieuan Runswick, Michael J. |
| Copyright Year | 1979 |
| Description | Journal: Journal of Biological Inorganic Chemistry 4-(3-Bromoacetylpyridinio)butyldiphosphoadenosine was synthesized with a [carbonyl-14C]acetyl label. The reactive coenzyme analogue inactivates alcohol dehydrogenase from Bacillus stearothermophilus by forming a covalent enzyme-coenzyme compound. The inactivation kinetics as well as the spectral properties of the modified enzyme after treatment with sodium hyposulphite suggest that the analogue is bound at the coenzyme binding site. B. stearothermophilus alcohol dehydrogenase modified with 14C-labelled coenzyme analogue and subseqeuntly carboxymethylated with unlabelled iodoacetic acid was digested with trypsin. The radioactive peptide was isolated and sequenced in parallel with the corresponding peptide similarly isolated from unmodified enzyme that had instead been carboxymethylated with iodo[14C]acetic acid. Amino acid and sequence analysis show that Cys-38 of the B. stearothermophilus alcohol dehydrogenase was modified by the reactive coenzyme analogue. This residue is homologous to Cys-43 in yeast alcohol dehydrogenase and Cys-46 in the horse liver enzyme but, unlike the latter two, Cys-38 is not reactive towards iodoacetate in the native bacterial enzyme. |
| Related Links | http://onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1979.tb12794.x/pdf |
| Ending Page | 64 |
| Page Count | 8 |
| Starting Page | 57 |
| ISSN | 09498257 |
| e-ISSN | 14321327 |
| DOI | 10.1111/j.1432-1033.1979.tb12794.x |
| Journal | Journal of Biological Inorganic Chemistry |
| Issue Number | 1 |
| Volume Number | 93 |
| Language | English |
| Publisher | Wiley-Blackwell |
| Publisher Date | 1979-01-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: Journal of Biological Inorganic Chemistry Biochemistry and Molecular Biology Stearothermophilus Bromoacetylpyridinio Butyldiphosphoadenosine |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry Inorganic Chemistry |
