title: Positions 94-98 of the lactose repressor N-subdomain monomer-monomer interface are critical for allosteric communication.
abstract: The central region of the LacI N-subdomain monomer-monomer interface includes residues K84, V94, V95, V96, S97, and M98. The side chains of these residues line the β-strands at this interface and interact to create a network of hydrophobic, charged, and polar interactions that significantly rearranges in different functional states of LacI. Prior work showed that converting K84 to an apolar residue or converting V96 to an acidic residue impedes the allosteric response to inducer. Thus, we postulated that a disproportionate number of substitutions in this region of the monomer-monomer interface would alter the complex features of the LacI allosteric response. To explore this hypothesis, acidic, basic, polar, and apolar mutations were introduced at positions 94-98. Despite their varied locations along the β-strands that flank the interface, ∼70% of the mutations impact allosteric behavior, with the most significant effects found for charged substitutions. Of note, many of the LacI variants with minor functional impact exhibited altered stability to urea denaturation. The results confirm the critical role of amino acids 94-98 and indicate that this N-subdomain interface forms a primary pathway in LacI allosteric response.
authors.affiliation: Department of Biochemistry and Cell Biology, Rice University, MS-140, 6100 South Main Street, Houston, Texas 77005, USA.
authors.forename: Hongli
authors.initials: H
authors.lastname: Zhan
authors.forename: Maricela
authors.initials: M
authors.lastname: Camargo
authors.forename: Kathleen S
authors.initials: KS
authors.lastname: Matthews
mesh_terms: D000494:Allosteric Regulation; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D056968:Lac Repressors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation; D011485:Protein Binding; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D058767:Protein Unfolding
chemical_list: D029968:Escherichia coli Proteins; D056968:Lac Repressors; C539064:LacI protein, E coli; D004247:DNA
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