Patterns of nonadditivity between pairs of stability mutations in staphylococcal nuclease.


Abstract

To identify interactions between amino acid positions in staphylococcal nuclease that affect its stability, a collection of 71 double-mutant forms was constructed from 22 previously characterized single mutants. These single mutations were assigned to three different classes on the basis of their m value [m = d(delta G)/d[GuHCl]], a parameter that has been correlated with energetically significant changes in the structure of the denatured state [Green et al. (1992) Biochemistry 31,5717-5728]. Several mutant pairs from five of the six possible double-mutant classes were analyzed by guanidine hydrochloride denaturation to determine the extent to which changes in stability (delta delta GH2O) and changes in the m value (delta mGuHCl) reflect the sum of the effects of the individual mutants. The differences between the values for delta delta GH2O and delta mGuHCl estimated on the assumption of additivity and those obtained by experiment, i.e., delta delta delta G and delta delta m, were calculated for each double-mutant protein. Surprisingly, a large majority of double mutants from four of the five classes exhibited positive values of delta delta delta G and delta delta m; i.e., they were more stable and displayed a higher sensitivity to GuHCl than predicted on the basis of additivity.(ABSTRACT TRUNCATED AT 250 WORDS) Study holds ProTherm entries: 2659, 2660, 2661, 2662, 2663, 2664, 2665, 2666, 2667, 2668, 2669, 2670, 2671, 2672, 2673, 2674, 2675, 2676, 2677, 2678, 2679, 2680, 2681, 2682, 2683, 2684, 2685, 2686, 2687, 2688, 2689, 2690, 2691, 2692, 2693, 2694, 2695, 2696, 2697, 2698, 2699, 2700, 2701, 2702, 2703, 2704, 2705, 2706, 2707, 2708, 2709, 2710, 2711, 2712, 2713, 2714, 2715, 2716, 2717, 2718, 2719, 2720, 2721, 2722, 2723, 2724, 2725, 2726, 2727, 2728, 2729, 2730, 2731, 2732, 2733, 2734, 2735, 2736, 2737, 2738, 2739, 2740, 2741 Extra Details: m values are relative to the wild type value (6.85 kcal/mol/M),which has been normalized to 1.00 staphylococcal nuclease; protein stability; double-mutants;,nonadditivity; single mutants

Submission Details

ID: 6oGwPprL3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:19 p.m.

Version: 1

Publication Details
Green SM;Shortle D,Biochemistry (1993) Patterns of nonadditivity between pairs of stability mutations in staphylococcal nuclease. PMID:8399139
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
4WRD 2014-10-23T00:00:00+0000 1.65 Crystal structure of Staphylcoccal nulease variant Delta+PHS V66E L125E at cryogenic temperature
2LKV 2011-10-21T00:00:00+0000 0 Staphylococcal Nuclease PHS variant
2M00 2012-10-14T00:00:00+0000 0 Solution structure of staphylococcal nuclease E43S mutant in the presence of ssDNA and Cd2+
2OXP 2007-02-20T00:00:00+0000 2.0 Crystal Structure of Staphylococcal Nuclease mutant V66D/P117G/H124L/S128A
3D4W 2008-05-15T00:00:00+0000 1.9 Crystal structure of Staphylococcal nuclease variant Delta+PHS A109R at cryogenic temperature
3D8G 2008-05-23T00:00:00+0000 1.99 Crystal structure of Staphylococcal nuclease variant Delta+PHS I72R at cryogenic temperature
3MVV 2010-05-04T00:00:00+0000 1.72 Crystal structure of Staphylococcal nuclease variant Delta+PHS F34A at cryogenic temperature
3QOJ 2011-02-10T00:00:00+0000 1.6 Cryogenic structure of Staphylococcal nuclease variant D+PHS/V23K
3QOL 2011-02-10T00:00:00+0000 1.9 Crystal structure of Staphylococcal nuclease variant D+PHS/V23E at pH 6 determined at 100 K
3R3O 2011-03-16T00:00:00+0000 1.9 Crystal structure of Staphylococcal nuclease variant Delta+PHS T62A at cryogenic temperature and with high redundancy

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
99.3 Thermonuclease Q6GIK1 NUC_STAAR
99.3 Thermonuclease Q8NXI6 NUC_STAAW
99.3 Thermonuclease Q6GB41 NUC_STAAS
99.1 Thermonuclease Q7A6P2 NUC_STAAN
99.1 Thermonuclease Q99VJ0 NUC_STAAM
99.3 Thermonuclease Q5HHM4 NUC_STAAC
100.0 Thermonuclease P00644 NUC_STAAU