Item request has been placed! ×
Item request cannot be made. ×
loading  Processing Request

Guanine-nucleotide binding and hydrolyzing kinetics of ORrab2, a rice small GTP-binding protein expressed in Escherichia coli.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
اقرأ على الانترنت اقرأ أكثر حفظ في قائمتي
  • المؤلفون: Seo HS;Seo HS; Choi CH; Kim HY; Jeong JY; Lee SY; Cho MJ; Bahk JD
  • المصدر:
    European journal of biochemistry [Eur J Biochem] 1997 Oct 01; Vol. 249 (1), pp. 293-300.
  • نوع النشر :
    Journal Article; Research Support, Non-U.S. Gov't
  • اللغة:
    English
  • معلومة اضافية
    • المصدر:
      Publisher: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies Country of Publication: England NLM ID: 0107600 Publication Model: Print Cited Medium: Print ISSN: 0014-2956 (Print) Linking ISSN: 00142956 NLM ISO Abbreviation: Eur J Biochem Subsets: MEDLINE
    • بيانات النشر:
      Publication: -2004: Oxford, UK : Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
      Original Publication: Berlin, New York, Springer.
    • الموضوع:
      rab GTP-Binding Proteins*; GTP-Binding Proteins/*metabolism ; Oryza/*metabolism ; Plant Proteins/*metabolism; Amino Acid Sequence ; Escherichia coli/genetics ; GTP Phosphohydrolases/genetics ; GTP Phosphohydrolases/metabolism ; GTP-Binding Proteins/genetics ; Guanine Nucleotides/metabolism ; Kinetics ; Molecular Sequence Data ; Oryza/genetics ; Plant Proteins/genetics ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Sequence Homology, Amino Acid
    • نبذة مختصرة :
      The ORrab2 gene encodes a GTP-binding protein of 23.169 kDa. The deduced amino acid sequence shows that ORrab2 has the motifs conserved among small GTP-binding proteins in plants and that it shares sequence identity with Atrab2 (93.0%), Hrab2 (85.2%), Hrab4 (51.9%), Hrab1 (46.2%), YPT (40.7%), Hrab3B (40.0%), Hrab3A (38.1%), SEC4 (38.1%), Hrab5 (34.3%) and Hrab6 (32.4%). To analyze the biochemical properties of this protein, an ORrab2 cDNA was overexpressed in Escherichia coli and the protein purified by Ni2+-nitrilotriacetic acid agarose and hydroxyapatite column chromatography. The molecular mass of the protein bearing a His-tag is approximately 28.2 kDa. The guanine-nucleotide binding and hydrolyzing activity of ORrab2 increased with non-ionic C12E10 (polyoxyethylene 10-lauryl ether) and ionic Chaps detergent treatment. ORrab2 bound maximally 1.03 mol of [gamma-35S]GTP[S]/mol of protein with a Kd value of 56.83 nM. The ratios k(off GDP)/k(off GTP) of ORrab2 were 3.63 for the control, 3.7 in the presence of C12E10, and 3.83 with Chaps, indicating that ORrab2 has a higher affinity for GTP than GDP. The rate (k(cat)) of Pi release against [gamma-32P]GTP bound ORrab2 in a steady state and the rate of hydrolysis of [gamma-32P]GTP (kGTPase) were calculated to be 432 x 10(-4) +/- 8 x 10(-4) min(-1) and 172 x 10(-4) +/- 2 x 10(-4) min(-1), respectively, in the presence of 0.1% C12E10 and 1 mM MgSO4.
    • الرقم المعرف:
      0 (Guanine Nucleotides)
      0 (Plant Proteins)
      0 (Recombinant Proteins)
      0 (rab2 protein, Oryza sativa)
      EC 3.6.1.- (GTP Phosphohydrolases)
      EC 3.6.1.- (GTP-Binding Proteins)
      EC 3.6.5.2 (rab GTP-Binding Proteins)
    • الموضوع:
      Date Created: 19971118 Date Completed: 19971205 Latest Revision: 20190620
    • الموضوع:
      20240829
    • الرقم المعرف:
      10.1111/j.1432-1033.1997.00293.x
    • الرقم المعرف:
      9363782