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CUREs for high-level Galectin-3 expression.
Charbonneau, Alexander A; Reicks, Elizabeth J; Cambria, John F; Inman, Jacob; Danley, Daria; Shockley, Emmie A; Davion, Ravenor; Salgado, Isabella; Norton, Erienne G; Corbett, Lucy J; Hanacek, Lucy E; Jensen, Jordan G; Kibodeaux, Marguerite A; Kirkpatrick, Tess K; Rausch, Keilen M; Roth, Samantha R; West, Bernadette; Wilson, Kenai E; Lawrence, C Martin; Cloninger, Mary J.
Afiliación
  • Charbonneau AA; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Reicks EJ; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Cambria JF; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Inman J; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Danley D; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Shockley EA; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Davion R; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Salgado I; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Norton EG; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Corbett LJ; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Hanacek LE; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Jensen JG; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Kibodeaux MA; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Kirkpatrick TK; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Rausch KM; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Roth SR; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • West B; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Wilson KE; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Lawrence CM; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA.
  • Cloninger MJ; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 59717, USA. Electronic address: mcloninger@montana.edu.
Protein Expr Purif ; 221: 106516, 2024 Sep.
Article en En | MEDLINE | ID: mdl-38801985
ABSTRACT
Galectins are a large and diverse protein family defined by the presence of a carbohydrate recognition domain (CRD) that binds ß-galactosides. They play important roles in early development, tissue regeneration, immune homeostasis, pathogen recognition, and cancer. In many cases, studies that examine galectin biology and the effect of manipulating galectins are aided by, or require the ability to express and purify, specific members of the galectin family. In many cases, E. coli is employed as a heterologous expression system, and galectin expression is induced with isopropyl ß-galactoside (IPTG). Here, we show that galectin-3 recognizes IPTG with micromolar affinity and that as IPTG induces expression, newly synthesized galectin can bind and sequester cytosolic IPTG, potentially repressing further expression. To circumvent this putative inhibitory feedback loop, we utilized an autoinduction protocol that lacks IPTG, leading to significantly increased yields of galectin-3. Much of this work was done within the context of a course-based undergraduate research experience, indicating the ease and reproducibility of the resulting expression and purification protocols.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Galectina 3 / Escherichia coli / Isopropil Tiogalactósido Límite: Humans Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Galectina 3 / Escherichia coli / Isopropil Tiogalactósido Límite: Humans Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos