Higher-order packing interactions in triple and quadruple mutants of staphylococcal nuclease.


Abstract

Sixty-four triple and 32 quadruple mutants were constructed in the core of staphylococcal nuclease. This is the first time that a large number of multiple mutants with all possible variations and all possible lower-order mutants has been systematically constructed in any protein core. Stabilities were determined by solvent denaturation. The energetic effects of these multiple mutants have been analyzed in combination with the stability data from the component single and double mutants. It was found that most of the stability changes in triple and quadruple mutants cannot be correctly predicted from stability effects of component single mutants. However, if the interaction energy between pairs of side chains in the component double mutants is taken into account, correct stability prediction can be made for most triple and quadruple mutants. The data further show that while packing interactions unique to triple and quadruple mutations do occur, they are of much less energetic significance than interactions between pairs of residues. The results presented here show that the packing of a protein interior can be closely approximated in most cases as a series of short-range, nearest-neighbor interactions. This has profound implications for rational protein design and structure prediction. Study holds ProTherm entries: 12481, 12482, 12483, 12484, 12485, 12486, 12487, 12488, 12489, 12490, 12491, 12492, 12493, 12494, 12495, 12496, 12497, 12498, 12499, 12500, 12501, 12502, 12503, 12504, 12505, 12506, 12507, 12508, 12509, 12510, 12511, 12512, 12513, 12514, 12515, 12516, 12517, 12518, 12519, 12520, 12521, 12522, 12523, 12524, 12525, 12526, 12527, 12528, 12529, 12530, 12531, 12532, 12533, 12534, 12535, 12536, 12537, 12538, 12539, 12540, 12541, 12542, 12543, 12544, 12545, 12546, 12547, 12548, 12549, 12550, 12551, 12552, 12553, 12554, 12555, 12556, 12557, 12558, 12559, 12560, 12561, 12562, 12563, 12564, 12565, 12566, 12567, 12568, 12569, 12570, 12571, 12572, 12573, 12574, 12575, 12576, 12577 Extra Details: quadruple mutants; core; interaction energy; packing interactions

Submission Details

ID: oC9QRndv

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:43 p.m.

Version: 1

Publication Details
Chen J;Stites WE,Biochemistry (2001) Higher-order packing interactions in triple and quadruple mutants of staphylococcal nuclease. PMID:11705393
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 UniProtKB Entries

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