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

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
2GVB 1995-10-15 REFINED SOLUTION STRUCTURE OF THE TYR 41--> HIS MUTANT OF THE M13 GENE V PROTEIN. A COMPARISON WITH THE CRYSTAL STRUCTURE
2GVA 1995-10-15 REFINED SOLUTION STRUCTURE OF THE TYR 41--> HIS MUTANT OF THE M13 GENE V PROTEIN. A COMPARISON WITH THE CRYSTAL STRUCTURE
1GVP 1997-09-04 1.6 GENE V PROTEIN (SINGLE-STRANDED DNA BINDING PROTEIN)
1GKH 1997-09-04 1.7 MUTANT K69H OF GENE V PROTEIN (SINGLE-STRANDED DNA BINDING PROTEIN)
1VQI 1997-02-12 1.8 GENE V PROTEIN MUTANT WITH ILE 47 REPLACED BY VAL 47 (I47V)
1VQB 1997-02-12 1.8 GENE V PROTEIN (SINGLE-STRANDED DNA BINDING PROTEIN)
1VQH 1997-02-12 1.8 GENE V PROTEIN MUTANT WITH ILE 47 REPLACED BY MET 47 (I47M)
1VQJ 1997-02-12 1.8 GENE V PROTEIN MUTANT WITH VAL 35 REPLACED BY ILE 35 (V35I)
1VQF 1997-02-12 1.8 GENE V PROTEIN MUTANT WITH VAL 35 REPLACED BY ILE 35 AND ILE 47 REPLACED BY VAL 47 (V35I, I47V)
1VQE 1997-02-12 1.8 GENE V PROTEIN MUTANT WITH VAL 35 REPLACED BY ILE 35 AND ILE 47 REPLACED BY MET 47 (V35I, I47M)
1VQC 1997-02-12 1.8 GENE V PROTEIN MUTANT WITH VAL 35 REPLACED BY ILE 35 AND ILE 47 REPLACED BY PHE 47 (V35I, I47F)
1VQA 1997-02-12 1.8 GENE V PROTEIN MUTANT WITH VAL 35 REPLACED BY ALA 35 AND ILE 47 REPLACED BY LEU 47 (V35A, I47L)
1VQD 1997-02-12 1.82 GENE V PROTEIN MUTANT WITH VAL 35 REPLACED BY ILE 35 AND ILE 47 REPLACED BY LEU 47 (V35I, I47L)
1VQG 1997-02-12 1.82 GENE V PROTEIN MUTANT WITH ILE 47 REPLACED BY LEU 47 (I47L)
1AE2 1997-09-04 2.0 MUTANT L32R OF GENE V PROTEIN (SINGLE-STRANDED DNA BINDING PROTEIN)
1AE3 1997-09-04 2.0 MUTANT R82C OF GENE V PROTEIN (SINGLE-STRANDED DNA BINDING PROTEIN)
1YHB 1994-06-22 2.2 CRYSTAL STRUCTURES OF Y41H AND Y41F MUTANTS OF GENE V PROTEIN FROM FF PHAGE SUGGEST POSSIBLE PROTEIN-PROTEIN INTERACTIONS IN GVP-SSDNA COMPLEX
2GN5 1986-01-21 2.3 REFINED STRUCTURE OF THE GENE 5 DNA BINDING PROTEIN FROM BACTERIOPHAGE FD
1YHA 1994-06-22 2.5 CRYSTAL STRUCTURES OF Y41H AND Y41F MUTANTS OF GENE V PROTEIN FROM FF PHAGE SUGGEST POSSIBLE PROTEIN-PROTEIN INTERACTIONS IN GVP-SSDNA COMPLEX

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 DNA-Binding protein G5P P69544 G5P_BPM13
100.0 DNA-Binding protein G5P P69542 G5P_BPFD
100.0 DNA-Binding protein G5P P69543 G5P_BPF1