Relevance of glycine and cysteine residues as well as N- and C-terminals for the activity of protein histidine phosphatase
| Autor: | Susanne Klumpp, Nicole Bäumer, Nien Tze Ma, Gunther Bechmann, Josef Krieglstein |
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| Rok vydání: | 2010 |
| Předmět: |
musculoskeletal diseases
Alanine chemistry.chemical_classification Chemistry Phosphatase Glycine Biophysics Lyase Biochemistry Phosphoric Monoester Hydrolases Analytical Chemistry Amino acid Escherichia coli Mutagenesis Site-Directed Humans natural sciences Amino Acid Sequence Cysteine Cloning Molecular Binding site Site-directed mutagenesis Molecular Biology Histidine |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 1804:206-211 |
| ISSN: | 1570-9639 |
| Popis: | There is increasing evidence that reversible phosphorylation of histidine residues regulates numerous important cellular processes. The first protein histidine phosphatase (PHP) from vertebrates was discovered just recently. Here, we report on amino acids and domains essential for activity of PHP. Point mutations of conserved residues and deletions of the N- and C-termini of PHP were analyzed using [ 32 P-his]ATP-citrate lyase as a substrate. Individual or joint replacement of all cysteine residues by alanine did not affect PHP activity. Deletion of 9 N-terminal amino acids resulted in inactive PHP. Furthermore, only 4 C-terminal residues could be deleted without losing PHP activity. Single or multiple mutations of the glycine-rich domain (Gly 75 , Gly 77 ) of a putative nucleotide binding site of PHP (GxGxxG/S) caused inactivation of PHP. Wildtype PHP could be labeled with [α- 32 P]ATP. Such radiolabeling was not detectable for catalytically inactive PHP-G75A and PHP-G77A. These data suggest further studies on the interaction between PHP and ATP. |
| Databáze: | OpenAIRE |
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