The tetrameric structure of nucleotide-regulated pyrophosphatase and its modulation by deletion mutagenesis and ligand binding
| Autor: | Alexander A. Baykov, Anu Salminen, Victor N. Orlov, Viktor A. Anashkin, Reijo Lahti |
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| Rok vydání: | 2020 |
| Předmět: |
inorganic chemicals
0301 basic medicine congenital hereditary and neonatal diseases and abnormalities Stereochemistry Biophysics CBS domain Desulfitobacterium Ligands Pyrophosphate Biochemistry Cofactor 03 medical and health sciences chemistry.chemical_compound Tetramer Bacterial Proteins Adenine nucleotide Catalytic Domain Nucleotide Amino Acid Sequence Molecular Biology Pyrophosphatases Sequence Deletion chemistry.chemical_classification Pyrophosphatase 030102 biochemistry & molecular biology biology organic chemicals nutritional and metabolic diseases Desulfitobacterium hafniense biology.organism_classification Inorganic Pyrophosphatase 030104 developmental biology chemistry Mutagenesis biology.protein |
| Zdroj: | Archives of biochemistry and biophysics. 692 |
| ISSN: | 1096-0384 |
| Popis: | A quarter of prokaryotic Family II inorganic pyrophosphatases (PPases) contain a regulatory insert comprised of two cystathionine β-synthase (CBS) domains and one DRTGG domain in addition to the two catalytic domains that form canonical Family II PPases. The CBS domain-containing PPases (CBS-PPases) are allosterically activated or inhibited by adenine nucleotides that cooperatively bind to the CBS domains. Here we use chemical cross-linking and analytical ultracentrifugation to show that CBS-PPases from Desulfitobacterium hafniense and four other bacterial species are active as 200–250-kDa homotetramers, which seems unprecedented among the four PPase families. The tetrameric structure is stabilized by Co2+, the essential cofactor, pyrophosphate, the substrate, and adenine nucleotides, including diadenosine tetraphosphate. The deletion variants of dhPPase containing only catalytic or regulatory domains are dimeric. Co2+ depletion by incubation with EDTA converts CBS-PPase into inactive tetrameric and dimeric forms. Dissociation of tetrameric CBS-PPase and its catalytic part by dilution renders them inactive. The structure of CBS-PPase tetramer was modelled from the structures of dimeric catalytic and regulatory parts. These findings signify the role of the unique oligomeric structure of CBS-PPase in its multifaced regulation. |
| Databáze: | OpenAIRE |
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