Understanding Thermal Adaptation of Enzymes through the Multistate Rational Design and Stability Prediction of 100 Adenylate Kinases S


Abstract

Careful balance between structural stability and flexibility is a hallmark of enzymatic function, and temperature can affect both properties. Canonical (fixed-backbone) enzyme design strategies currently do not consider the role of these properties. Herein, we describe the rational design of 100 temperature-adapted adenylate kinase enzymes using a multistate design strategy that incorporates the impact of conformational changes to backbone structure and stability, in addition to experimental analysis of thermostability and function. Comparison of the experimental temperature of maximum activity to the melting temperature across all 100 variants reveals a strong correlation between these two parameters. In turn, experimental stability data were used to produce accurate predictions of thermostability, providing the requisite complement for de novo temperature-adapted enzyme design. In principle, this level of design-based analysis can be applied to any protein, paving the way toward identifying and understanding the hallmarks of the thermodynamic and structural limits of function.

Submission Details

ID: zeSyKHyk

Submitter: Barry Olafson

Submission Date: July 31, 2017, 11:46 a.m.

Version: 1

Publication Details
Howell SC;Inampudi KK;Bean DP;Wilson CJ,Structure (2014) Understanding thermal adaptation of enzymes through the multistate rational design and stability prediction of 100 adenylate kinases. PMID:24361272
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
1P3J 2003-04-17T00:00:00+0000 1.9 Adenylate Kinase from Bacillus subtilis
2EU8 2005-10-28T00:00:00+0000 1.8 Crystal structure of a thermostable mutant of Bacillus subtilis Adenylate Kinase (Q199R)
2OO7 2007-01-25T00:00:00+0000 1.8 Crystal structure of a thermostable mutant of Bacillus subtilis Adenylate Kinase (T179I/Q199R)
2ORI 2007-02-02T00:00:00+0000 1.8 Crystal structure of a thermostable mutant of Bacillus subtilis Adenylate Kinase (A193V/Q199R/)
2OSB 2007-02-05T00:00:00+0000 1.8 Crystal structure of a thermostable mutant of Bacillus subtilis Adenylate Kinase (Q16L/Q199R/)
2P3S 2007-03-09T00:00:00+0000 1.8 Crystal structure of a thermostable mutant of Bacillus subtilis Adenylate Kinase (G214R/Q199R)
2QAJ 2007-06-15T00:00:00+0000 1.8 Crystal structure of a thermostable mutant of Bacillus subtilis Adenylate Kinase (Q199R/G213E)
3DKV 2008-06-26T00:00:00+0000 1.82 Crystal structure of adenylate kinase variant AKlse1
3DL0 2008-06-26T00:00:00+0000 1.58 Crystal structure of adenylate kinase variant AKlse3
4MKF 2013-09-05T00:00:00+0000 1.7 Crystal structure of a stable adenylate kinase variant AKv3

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Adenylate Kinase P16304 KAD_BACSU
95.4 Adenylate Kinase A7Z0Q9 KAD_BACVZ