Structure of the ribosome associating GTPase HflX
Hao Wu
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Protein Studies Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma
Hao Wu, Lei Sun, and Fabian Blombach contributed equally to this work.
Search for more papers by this authorLei Sun
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Hao Wu, Lei Sun, and Fabian Blombach contributed equally to this work.
Search for more papers by this authorFabian Blombach
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
Hao Wu, Lei Sun, and Fabian Blombach contributed equally to this work.
Search for more papers by this authorStan J.J. Brouns
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
Search for more papers by this authorAmbrosius P. L. Snijders
Department of Chemical and Process Engineering, University of Sheffield, Sheffield S1 3JD, UK
Search for more papers by this authorKristina Lorenzen
Department of Biomolecular Mass Spectrometry, Utrecht University, Utrecht, The Netherlands
Search for more papers by this authorRobert H. H. van den Heuvel
Department of Biomolecular Mass Spectrometry, Utrecht University, Utrecht, The Netherlands
Search for more papers by this authorAlbert J. R. Heck
Department of Biomolecular Mass Spectrometry, Utrecht University, Utrecht, The Netherlands
Search for more papers by this authorSheng Fu
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorXuemei Li
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorXuejun C. Zhang
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Protein Studies Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma
Search for more papers by this authorZihe Rao
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorCorresponding Author
John van der Oost
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, The Netherlands===Search for more papers by this authorHao Wu
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Protein Studies Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma
Hao Wu, Lei Sun, and Fabian Blombach contributed equally to this work.
Search for more papers by this authorLei Sun
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Hao Wu, Lei Sun, and Fabian Blombach contributed equally to this work.
Search for more papers by this authorFabian Blombach
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
Hao Wu, Lei Sun, and Fabian Blombach contributed equally to this work.
Search for more papers by this authorStan J.J. Brouns
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
Search for more papers by this authorAmbrosius P. L. Snijders
Department of Chemical and Process Engineering, University of Sheffield, Sheffield S1 3JD, UK
Search for more papers by this authorKristina Lorenzen
Department of Biomolecular Mass Spectrometry, Utrecht University, Utrecht, The Netherlands
Search for more papers by this authorRobert H. H. van den Heuvel
Department of Biomolecular Mass Spectrometry, Utrecht University, Utrecht, The Netherlands
Search for more papers by this authorAlbert J. R. Heck
Department of Biomolecular Mass Spectrometry, Utrecht University, Utrecht, The Netherlands
Search for more papers by this authorSheng Fu
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorXuemei Li
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorXuejun C. Zhang
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Protein Studies Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma
Search for more papers by this authorZihe Rao
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorCorresponding Author
John van der Oost
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands
Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, The Netherlands===Search for more papers by this authorAbstract
The HflX-family is a widely distributed but poorly characterized family of translation factor-related guanosine triphosphatases (GTPases) that interact with the large ribosomal subunit. This study describes the crystal structure of HflX from Sulfolobus solfataricus solved to 2.0-Å resolution in apo- and GDP-bound forms. The enzyme displays a two-domain architecture with a novel “HflX domain” at the N-terminus, and a classical G-domain at the C-terminus. The HflX domain is composed of a four-stranded parallel β-sheet flanked by two α-helices on either side, and an anti-parallel coiled coil of two long α-helices that lead to the G-domain. The cleft between the two domains accommodates the nucleotide binding site as well as the switch II region, which mediates interactions between the two domains. Conformational changes of the switch regions are therefore anticipated to reposition the HflX-domain upon GTP-binding. Slow GTPase activity has been confirmed, with an HflX domain deletion mutant exhibiting a 24-fold enhanced turnover rate, suggesting a regulatory role for the HflX domain. The conserved positively charged surface patches of the HflX-domain may mediate interaction with the large ribosomal subunit. The present study provides a structural basis to uncover the functional role of this GTPases family whose function is largely unknown. Proteins 2010. © 2009 Wiley-Liss, Inc.
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