2Z1G

Crystal structure of E.coli RNase HI surface charged mutant(Q4R/T40E/Q72H/Q76K/Q80E/T92K/Q105K)

Summary for 2Z1G

• Entry DOI: 10.2210/pdb2z1g/pdb
• Related: 2Z1H 2Z1I 2Z1J
• Descriptor: Ribonuclease HI (2 entities in total)
• Functional Keywords: rnase hi, thermostability, surface-charge residue, hydrolase
• Biological source: Escherichia coli
• Cellular location: Cytoplasm (Potential): P0A7Y4
• Total number of polymer chains: 1
• Total formula weight: 17721.26

Authors

You, D.J.,Fukuchi, S.,Nishikawa, K.,Koga, Y.,Takano, K.,Kanaya, S. (deposition date: 2007-05-08, release date: 2007-11-13, Last modification date: 2023-11-01)

Primary citation

You, D.-J.,Fukuchi, S.,Nishikawa, K.,Koga, Y.,Takano, K.,Kanaya, S.
Protein Thermostabilization Requires a Fine-tuned Placement of Surface-charged Residues
J.Biochem.(Tokyo), 142:507-516, 2007

PubMed Abstract: Using the information from the genome projects, recent comparative studies of thermostable proteins have revealed a certain trend of amino acid composition in which polar residues are scarce and charged residues are rich on the protein surface. To clarify experimentally the effect of the amino acid composition of surface residues on the thermostability of Escherichia coli Ribonuclease HI (RNase HI), we constructed six variants in which five to eleven polar residues were replaced by charged residues (5C, 7Ca, 7Cb, 9Ca, 9Cb and 11C). The thermal denaturation experiments indicated that all of the variant proteins are 3.2-10.1 degrees C in Tm less stable than the wild proteins. The crystal structures of resultant protein variants 7Ca, 7Cb, 9Ca and 11C closely resemble that of E. coli RNase HI in their global fold, and several different hydrogen bonding and ion-pair interactions are formed by the mutations. Comparison of the crystal structures of these variant proteins with that of E. coli RNase HI reveals that thermal destabilization is apparently related to electrostatic repulsion of the charged residues with neighbours. This result suggests that charged residues of natural thermostable proteins are strictly posted on the surface with optimal interactions and without repulsive interactions.

PubMed: 17761696
DOI: 10.1093/jb/mvm157

Experimental method

X-RAY DIFFRACTION (2.1 Å)