Amino acid substitutions that increase the thermal stability of the lambda Cro protein.


Abstract

A mutant Cro protein, which bears the Ile-30----Leu substitution, is thermally unstable and degraded more rapidly than wild-type Cro in vivo. Using an antibody screen, we have isolated five different second site suppressor substitutions that reduce the proteolytic hypersensitivity of this mutant Cro protein. Two of the suppressor substitutions increase the thermal stability of Cro by 12 degrees C to 14 degrees C. These amino acid substitutions affect residues 16 and 26, which are substantially exposed to solvent in the crystal structure of wild-type Cro. Study holds ProTherm entries: 827, 828, 829, 830, 13437, 13438, 13439 Extra Details: enhanced stability; lambda Cro, genetic suppression;,intracellular proteolysis, antibody screen

Submission Details

ID: 2HjJmcCi3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:16 p.m.

Version: 1

Publication Details
Pakula AA;Sauer RT,Proteins (1989) Amino acid substitutions that increase the thermal stability of the lambda Cro protein. PMID:2780540
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
2A63 2006-06-06 Solution structure of a stably monomeric mutant of lambda Cro produced by substitutions in the ball-and-socket interface
1COP 1995-10-15 THREE-DIMENSIONAL DIMER STRUCTURE OF THE LAMBDA-CRO REPRESSOR IN SOLUTION AS DETERMINED BY HETERONUCLEAR MULTIDIMENSIONAL NMR
2ORC 1998-05-27 CRO REPRESSOR INSERTION MUTANT K56-[DGEVK], NMR, 32 STRUCTURES
2OVG 2008-01-08 1.35 Lambda Cro Q27P/A29S/K32Q triple mutant at 1.35 A in space group P3221
2ECS 2008-01-08 1.4 Lambda Cro mutant Q27P/A29S/K32Q at 1.4 A in space group C2
1ORC 1996-12-23 1.54 CRO REPRESSOR INSERTION MUTANT K56-[DGEVK]
1D1M 1999-09-24 2.05 CRYSTAL STRUCTURE OF CRO K56-[DGEVK]-F58W MUTANT
1D1L 1999-10-06 2.1 CRYSTAL STRUCTURE OF CRO-F58W MUTANT
5CRO 1998-06-17 2.3 REFINED STRUCTURE OF CRO REPRESSOR PROTEIN FROM BACTERIOPHAGE LAMBDA
3ORC 1998-12-02 3.0 CRYSTAL STRUCTURE OF AN ENGINEERED CRO MONOMER BOUND NONSPECIFICALLY TO DNA
6CRO 1998-09-18 3.0 CRYSTAL STRUCTURE OF LAMBDA-CRO BOUND TO A CONSENSUS OPERATOR AT 3.0 ANGSTROM RESOLUTION
4CRO 1992-01-15 3.9 PROTEIN-DNA CONFORMATIONAL CHANGES IN THE CRYSTAL STRUCTURE OF A LAMBDA CRO-OPERATOR COMPLEX

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Regulatory protein cro P03040 RCRO_LAMBD