1JYE

Structure of a Dimeric Lac Repressor with C-terminal Deletion and K84L Substitution

Summary for 1JYE

• Entry DOI: 10.2210/pdb1jye/pdb
• Related: 1JYF
• Descriptor: Lactose Operon Repressor, GLYCEROL (3 entities in total)
• Functional Keywords: gene regulation, protein stability, protein dna-binding, transcription
• Biological source: Escherichia coli
• Total number of polymer chains: 1
• Total formula weight: 37518.78

Authors

Bell, C.E.,Barry, J.,Matthews, K.S.,Lewis, M. (deposition date: 2001-09-12, release date: 2001-10-18, Last modification date: 2024-04-03)

Primary citation

Bell, C.E.,Barry, J.,Matthews, K.S.,Lewis, M.
Structure of a variant of lac repressor with increased thermostability and decreased affinity for operator.
J.Mol.Biol., 313:99-109, 2001

PubMed Abstract: A single amino acid substitution, K84L, in the Escherichia coli lac repressor produces a protein that has substantially increased stability compared to wild-type. However, despite the increased stability, this altered tetrameric repressor has a tenfold reduced affinity for operator and greatly decreased rate-constants of inducer binding as well as a reduced phenotypic response to inducer in vivo. To understand the dramatic increase in stability and altered functional properties, we have determined the X-ray crystal structures of a dimeric repressor with and without the K84L substitution at resolutions of 1.7 and 3.0 A, respectively. In the wild-type dimer, K84-11, Lys84 forms electrostatic interactions at the monomer-monomer interface and is partially exposed to solvent. In the K84L-11 substituted protein there is reorientation of the N-subdomains, which allows the leucine to become deeply buried at the monomer-monomer interface. This reorientation of the N-subdomains, in turn, results in an alteration of hydrogen bonding, ion pairing, and van der Waals interactions at the monomer-monomer interface. The lysine residue at position 84 appears to exert its key effects by destabilizing the "optimal" conformation of the repressor, effectively loosening the dimer interface and allowing the repressor to adopt the conformations necessary to function as a molecular switch.

PubMed: 11601849
DOI: 10.1006/jmbi.2001.5041

Experimental method

X-RAY DIFFRACTION (1.7 Å)