On the Interaction of Ribosomal Protein L5 with 5S rRNA(Biochemistry & Molecular Biology)

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概要

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• Ribosomal protein L5, a 5S rRNA binding protein in the large subunit, is composed of a five-stranded antiparallel β-sheet and four α-helices, and folds in a way that is topologically similar to the ribonucleprotein (RNP) domain [Nakashima et al., RNA 7, 692-701, 2001]. The crystal structure of ribosomal protein L5 (BstL5) from Bacillus stearothermophilus suggests that a concave surface formed by an anti-parallel β-sheet and long loop structures are strongly involved in 5S rRNA binding. To identify amino acid residues responsible for 5S rRNA binding, we made use of Ala-scanning mutagenesis of evolutionarily conserved amino acids occurred at β-strands and loop structures in BstL5. The mutation of Lys33 at the β1-strand caused a significant reduction in 5S rRNA binding. In addition, the Arg92, Phe122, and Glu134 mutations on the β2-strand, the α3-β4 loop, and the β4-β5 loop, respectively, resulted in a moderate decrease in the 5S rRNA binding affinity. In contrast, mutation of the conserved residue Pro65 at the β2-strand had little effect on the 5S rRNA binding activity. These results, taken together with previous results, identified Lys33, Asn37, Gln63, and Thr90 on the β-sheet structure, and Phe77 at the β2-β3 loop as critical residues for the 5S rRNA binding. The contribution of these amino acids to 5S rRNA binding was further quantitatively evaluated by surface plasmon resonance (SPR) analysis by the use of BIAcore. The results showed that the amino acids on the β-sheet structure are required to decrease the dissociation rate constant for the BstL5-5S rRNA complex, while those on the loops are to increase the association rate constant for the BstL5-5S rRNA interaction.

• 社団法人日本農芸化学会の論文
• 2002-01-23

著者

• Kawamura Shunsuke

  東北薬科大学 医療薬学

• Kawamura Shunsuke

  Laboratory Of Biochemistry Faculty Of Agriculture Kyushu University

• Kawamura Shunsuke

  Laboratorv Of Cell Biology. School Of Agriculture' Kvushu Tokai University

• KIMURA Makoto

  Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University

• KIKUKAWA Shingo

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Grad

• TANAKA Isao

  Division of Biological Science, Graduate School of Science, Hokkaido University

• KOUZUMA Yoshiaki

  Laboratory of Biochemistry, Faculty of Agriculture, Graduate School, Kyushu University

• Kimura M

  Laboratory Of Biochemistry Faculty Of Agriculture Graduate School Kyushu University

• Tanaka I

  Division Of Biological Science Graduate School Of Science Hokkaido University

• IWASAKI Kenta

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Grad

• Kawamura S

  Department Of Physiology Keio University School Of Medicine

• Kouzuma Yoshiaki

  Laboratory Of Food Molecular Functionality College Of Agriculture Ibaraki University

• Kouzuma Yoshiaki

  Laboratory Of Food Functionality School Of Agriculture Ibaraki University

• Kawabe Masato

  United Graduate School Of Agricultural Science Tokyo University Of Agriculture And Technology (tuat)

• Iwasaki K

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat

• Iwasaki Kenta

  Laboratory Of Biochemistry Division Of Bioresource And Bioenvironmental Sciences Graduate School Of

• Tanaka I

  Fukui Prefecture Agricultural Experiment Station Fukui Jpn

• Kikukawa Shingo

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Graduate School Faculty Of Agr

• Kimura Makoto

  Digital Human Research Center Aist Japan

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Agricultural Chemistry Faculty And Graduate School Of Agric

• KOUZUMA Yoshiaki

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University

• KIMURA Makoto

  Laboratory for Remediation Research, Plant Science Center, RIKEN

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