Adenylyl cyclase Rv1625c of Mycobacterium tuberculosis: a progenitor of mammalian adenylyl cyclases
| Autor: | Thomas Seebacher, Ursula Kurz, Joachim E. Schultz, Ying Lan Guo, Jürgen U. Linder |
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| Rok vydání: | 2001 |
| Předmět: |
Gs alpha subunit
Molecular Sequence Data Biology Cyclase Article General Biochemistry Genetics and Molecular Biology ADCY10 Cell Line Evolution Molecular Adenylyl cyclase chemistry.chemical_compound Escherichia coli Humans Amino Acid Sequence Molecular Biology ADCY6 Sequence Homology Amino Acid General Immunology and Microbiology General Neuroscience ADCY9 Membrane Proteins Mycobacterium tuberculosis ADCY3 Recombinant Proteins Amino Acid Substitution chemistry Biochemistry cAMP-dependent pathway Dimerization Adenylyl Cyclases |
| Zdroj: | The EMBO Journal. 20:3667-3675 |
| ISSN: | 1460-2075 |
| Popis: | The gene Rv1625c from Mycobacterium tuberculosis encodes a membrane-anchored adenylyl cyclase corresponding to exactly one-half of a mammalian adenylyl cyclase. An engineered, soluble form of Rv1625c was expressed in Escherichia coli. It formed a homodimeric cyclase with two catalytic centers. Amino acid mutations predicted to affect catalysis resulted in inactive monomers. A single catalytic center with wild-type activity could be reconstituted from mutated monomers in stringent analogy to the mammalian heterodimeric cyclase structure. The proposed existence of supramolecular adenylyl cyclase complexes was established by reconstitution from peptide-linked, mutation-inactivated homodimers resulting in pseudo-trimeric and -tetrameric complexes. The mycobacterial holoenzyme was expressed successfully in E.coli and mammalian HEK293 cells, i.e. its membrane targeting sequence was compatible with the bacterial and eukaryotic machinery for processing and membrane insertion. The membrane-anchored mycobacterial cyclase expressed in E.coli was purified to homogeneity as a first step toward the complete structural elucidation of this important protein. As the closest progenitor of the mammalian adenylyl cyclase family to date, the mycobacterial cyclase probably was spread by horizontal gene transfer. |
| Databáze: | OpenAIRE |
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