Guanosine 5'-diphosphate 3'-diphosphate (ppGpp) Synthetic Activities on Escherichia coli SpoT Domains(Biochemistry & Molecular Biology)

スポンサーリンク

概要

• 論文の詳細を見る

• Escherichia coli SpoT protein, with 702 amino acid residues, is a bifunctional enzyme catalyzing both guanosine 5'-diphosphate 3'-diphosphate (ppGpp) degradation and its synthesis. First, we investigated how many domains are included in SpoT protein, by limited hydrolysis of the protein with serine proteases, α-chymotrypsin, and elastase. Based on the results, we deduced that SpoT protein is composed of two major domains, an N-terminal half domain from Met1 to Phe373 and a C-terminal half domain from Glu374 to Asn702 (C-terminal end). In addition, by a further α-chymotrypsin digestion, two cleaved sites were found at Arg196 in the N-terminal half domain (D12) and at Lys475 in the C-terminal half domain (D34), to produce four minor domains, D1, D2, D3, and D4. Next, plasmids expressing the two major domains (D12 and D34) and four minor domains (D1, D2, D3, and D4) were constructed. Consequently, the deduced SpoT minor domains as well as the major domains were expressed as stable protein units, except for D4. D4 may also be folded into a stable protein in E. coli cells, since high expression of D4 from a plasmid results in host cell lethality. E. coli relA^-, spoT^- double null strains expressing D1, D2, and D12 recovered cell growth in M9 minimal medium, but the transformants of D3, D4, and D34 did not grow in the minimal medium. This indicates that ppGpp synthetic activities could be restricted in the N-terminal half domain (D12, D1, and D2).

• 社団法人日本農芸化学会の論文
• 2002-07-23

著者

• IWAMOTO Ryoko

  Department of Chemistry, Faculty of Science, Nara Women's University

• Iwamoto Ryoko

  Department Of Chemistry Faculty Of Science Nara Women's University

• Nishimura Akiko

  National Institute For Genetics Of Japan

• FUJITA Cchizuko

  Department of Chemistry, Faculty of Science, Nara Women's University

• IKEHARA Kenji

  Department of Chemistry, Faculty of Science, Nara Women's University

• Ikehara Kenji

  Department Of Chemistry Faculty Of Science

• Fujita Cchizuko

  Department Of Chemistry Faculty Of Science Nara Women's University

• FUJITA Chizuko

  Department of Chemistry, Faculty of Science, Nara Women's University

• Fujita Chizuko

  Department of Chemistry, Faculty of Science, Nara Women's University

関連論文

• Purification and Characterization of D-Glucosaminitol Dehydrogenase from Agrobacterium radiobacter
• Guanosine 5'-diphosphate 3'-diphosphate (ppGpp) Synthetic Activities on Escherichia coli SpoT Domains(Biochemistry & Molecular Biology)
• NUCLEASE ACTIVITY OF [GADV]-PEPTIDES : from "[GADV]-protein world" to "RNA-protein world"(From Molecules to Life-Like Systems & RNA World, International Symposium on Origins of Life and Astrobiology)
• A POSSIBLE EVOLUTIONARY PATHWAY OF THE GENETIC CODE DEDUCED FROM THE SNS HYPOTHESIS
• Identification of an Indispensable Amino Acid for ppGpp Synthesis of Escherichia coli SpoT Protein(Biochemistry & Molecular Biology)
• Contribution to Catalysis and to Stability of the Essential Cysteine Residue at the Active Site of D-Glucosaminate Dehydratase from Pseudomonas fluorescens
• Purification and Characterization of D-Glucosaminate Dehydratase from Pseudomonas fluorescens(Biological Chemistry)
• D-Glucosaminate Aldolase Activity of D-Glucosaminate Dehydratase from Pseudomonas fluorescens and Its Requirement for Mn^ Ion
• SNS HYPOTHESIS ON THE ORIGIN OF THE GENETIC CODE
• 遺伝子進化のシミュレーション--遺伝子の起原に関するGC-NSF(a)仮説の証拠
• The [GADV]-protein world hypothesis on the origin of life(Chemical Evolution, International Symposium on Origins of Life and Astrobiology)
• Mn^ in D-Glucosaminate Dehydratase from Pseudomonas fluorescens

もっと見る 閉じる

スポンサーリンク