2. Academic Validation
  3. Protocol for reconstituting enzymatic activities for ultra-large histone methyltransferases NSD1 and SETD2 using a baculovirus expression system

Protocol for reconstituting enzymatic activities for ultra-large histone methyltransferases NSD1 and SETD2 using a baculovirus expression system

  • STAR Protoc. 2025 Jul 18;6(3):103963. doi: 10.1016/j.xpro.2025.103963.
Chen-I Hsu 1,  Emily Yeh 2,  Carol C L Chen 3,  Mahnue Sahn 1,  Jia-Ray Yu 4
Affiliations

Affiliations

  • 1 Virginia Tech Fralin Biomedical Research Institute Cancer Research Center DC, Washington, DC 20012, USA.
  • 2 Department of Biochemistry & Molecular Biology, Faculty of Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
  • 3 Department of Biochemistry & Molecular Biology, Faculty of Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, Canada; Terry Fox Laboratory, British Columbia Cancer Research Institute, Vancouver, BC V5Z 1L3, Canada.
  • 4 Virginia Tech Fralin Biomedical Research Institute Cancer Research Center DC, Washington, DC 20012, USA. Electronic address: jiarayyu@gmail.com.
PMID: 40684435 DOI: 10.1016/j.xpro.2025.103963
Abstract

Expression and reconstitution of large proteins in the human proteome are challenging and often require the use of a mammalian cell expression system that is costly and inefficient. Here, we present a protocol to reconstitute catalytically active, full-length NSD1 (nuclear receptor binding SET domain protein 1) and SETD2 (SET domain containing 2, histone lysine methyltransferase) using a clonal baculovirus expression system. We describe steps for producing baculovirus, clonal selection, and protein reconstitution. We then detail procedures for using animatic assays. For complete details on the use and execution of this protocol, please refer to Hsu et al.1.

Keywords

Cell culture; Protein Biochemistry; Protein expression and purification.

Figures
Products