The role of phenylalanine 31 in maintaining the conformational stability of ribonuclease P2 from Sulfolobus solfataricus under extreme conditions of temperature and pressure.


Abstract

Ribonuclease P2 from the thermophilic archaebacterium Sulfolobus solfataricus is a small protein (7 kDa) with a known three-dimensional structure. Inspection of the structure and molecular dynamics simulation reveal that three aromatic residues (Phe5, Phe31, and Tyr33) from the hydrophobic core have a strong van der Waals interaction energy. We studied the thermodynamics of the heat, cold, and pressure-induced protein conformational changes of the wild type and of the F31A and F31Y mutants by analyzing the protein UV absorbance in the fourth derivative mode. The wild-type protein was extremely stable under all conditions of temperature and pressure. Heat and cold denaturation of both mutants, as well as denaturation by pressure of the F31A mutant, led to significant blue shifts of the derivative spectrum, indicating increased solvent exposure of Tyr33. For the F31Y mutant, high pressure (400 MPa) protected the protein against thermal denaturation. This study, probing the properties of the hydrophobic aromatic core, complements a thermal unfolding study which probes the overall structural changes [Knapp, S., Karshikoff, A., Berndt, K. D., Christova, P., Atanasov, B., & Ladenstein, R. (1996) J. Mol. Biol. 264,1132-1144]. The differences observed in response to extremes of temperature, pressure, and pH may be rationalized by an unfolding mechanism involving larger parts of the peripheral protein while the integrity of the hydrophobic core is maintained. Study holds ProTherm entries: 8800, 8801 Extra Details: hydrophobic core; van der Waals interaction energy; pressure-induced,protein conformational changes; thermal unfolding; structural changes

Submission Details

ID: aucnx3Ps3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:36 p.m.

Version: 1

Publication Details
Mombelli E;Afshar M;Fusi P;Mariani M;Tortora P;Connelly JP;Lange R,Biochemistry (1997) The role of phenylalanine 31 in maintaining the conformational stability of ribonuclease P2 from Sulfolobus solfataricus under extreme conditions of temperature and pressure. PMID:9220960
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1B4O 2000-01-05 NMR STUDY OF SSO7D MUTANT (F31A) MINIMIZED AVERAGE STRUCTURE
1JIC 1998-10-14 SOLUTION NMR STRUCTURE OF RECOMBINANT SSO7D WITH RNASE ACTIVITY, MINIMIZED AVERAGE STRUCTURE
1BBX 1998-10-14 NON-SPECIFIC PROTEIN-DNA INTERACTIONS IN THE SSO7D-DNA COMPLEX, NMR, 1 STRUCTURE
1SSO 1995-05-08 SOLUTION STRUCTURE AND DNA-BINDING PROPERTIES OF A THERMOSTABLE PROTEIN FROM THE ARCHAEON SULFOLOBUS SOLFATARICUS
2CVR 2006-08-29 NMR solution structure of sso7d mutant, K12L, 12 conformers
1C8C 2001-05-04 1.45 CRYSTAL STRUCTURES OF THE CHROMOSOMAL PROTEINS SSO7D/SAC7D BOUND TO DNA CONTAINING T-G MISMATCHED BASE PAIRS
1BNZ 1998-11-11 2.0 SSO7D HYPERTHERMOPHILE PROTEIN/DNA COMPLEX
5B02 2016-03-23 2.21 Structure of the prenyltransferase MoeN5 with a fusion protein tag of Sso7d
6J8V 2019-04-17 2.23 Structure of MOEN5-SSO7D fusion protein in complex with ligand 2
5B0I 2016-03-23 2.26 Structure of MoeN5-Sso7d fusion protein in complex with beta-octyl glucoside
5GWW 2017-09-20 2.3 Structure of MoeN5-Sso7d fusion protein in complex with a permethylated substrate analogue
6J8W 2019-04-17 2.35 Structure of MOEN5-SSO7D fusion protein in complex with lig 1
5GWV 2017-09-20 2.4 Structure of MoeN5-Sso7d fusion protein in complex with a substrate analogue
5B0J 2016-03-23 2.5 Structure of MoeN5-Sso7d fusion protein in complex with beta-undecyl maltoside
5B03 2016-03-23 2.6 Structure of MoeN5-Sso7d fusion protein in complex with geranyl pyrophosphate
5B0K 2016-03-23 2.75 Structure of MoeN5-Sso7d fusion protein in complex with beta-decyl maltoside
5B0L 2016-03-23 2.8 Structure of MoeN5-Sso7d fusion protein in complex with beta-nonyl glucoside
5B0M 2016-03-23 3.05 Structure of MoeN5-Sso7d fusion protein in complex with beta-dodecyl maltoside
5U1C 2017-01-11 3.9 Structure of tetrameric HIV-1 Strand Transfer Complex Intasome

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
93.2 DNA-binding protein 7a Q96X56 DN7_SULTO
96.7 DNA-binding protein 7a O59632 DN7B_SACSH
98.4 DNA-binding protein 7a F0NJT3 DN7B_SULIH
100.0 DNA-binding protein 7a D2PHL8 DN7A_SULID
98.4 DNA-binding protein 7a P39476 DN7D_SACS2
100.0 DNA-binding protein 7a P61990 DN7A_SACSH
100.0 DNA-binding protein 7a P61991 DN7A_SACS2