Details

Autor(en) / Beteiligte

• 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.

Titel

CUREs for high-level Galectin-3 expression

Ist Teil von

• Protein expression and purification, 2024-09, Vol.221, p.106516, Article 106516

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2024

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Galectins are a large and diverse protein family defined by the presence of a carbohydrate recognition domain (CRD) that binds β-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 β-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. •Full-length galectin-3 and its carbohydrate recognition domain were obtained.•High yields and purities were achieved using an autoinduction protocol.•Autoinduction afforded significantly higher yields than IPTG induction.