Engineering multiple properties of a protein by combinatorial mutagenesis.

Abstract

A method for simultaneously engineering multiple properties of a protein, based on the observed additivity of effects of individual mutations, is presented. We show that, for the gene V protein of bacteriophage f1, effects of double mutations on both protein stability and DNA binding affinity are approximately equal to the sums of the effects of the constituent single mutations. This additivity of effects implies that it is possible to deliberately construct mutant proteins optimized for multiple properties by combination of appropriate single mutations chosen from a characterized library. Study holds ProTherm entries: 543, 544, 545, 546, 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571, 572, 2228, 2229, 2230, 2231, 2232, 2233, 2234, 2235, 2236, 2237, 2238, 2239, 2240, 2241, 2242, 2243, 2244, 2245, 2246, 2247, 2248, 2249, 2250, 2251, 2252, 2253, 2254, 2255, 2256, 2257, 2258 Extra Details: additive : EDTA(1 mM),ddG values were measured in the presence of 2M GdnHCl gene V protein, bacteriophage f1, combinatorial mutagenesis;,protein stability; DNA binding affinity

Submission Details

ID: txzyHZBj

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:15 p.m.

Version: 1

Publication Details
Sandberg WS;Terwilliger TC,Proc. Natl. Acad. Sci. U.S.A. (1993) Engineering multiple properties of a protein by combinatorial mutagenesis. PMID:8378307
Additional Information

Study Summary

Number of data points 61
Proteins DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P ; DNA-Binding protein G5P
Unique complexes 61
Assays/Quantities/Protocols Experimental Assay: ddG ; Experimental Assay: dG_H2O
Libraries Mutations for sequence MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLVKITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK

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 DNA-Binding protein G5P P69543 G5P_BPF1
100.0 DNA-Binding protein G5P P69542 G5P_BPFD
100.0 DNA-Binding protein G5P P69544 G5P_BPM13