Guan-Da Syu1,2,3, Eric Johansen4, Heng Zhu4,5,6. 1. Department of Biotechnology and Bioindustry Sciences, National Cheng Kung University, Tainan, Taiwan, Republic of China. 2. International Center for Wound Repair and Regeneration, National Cheng Kung University, Tainan, Taiwan, Republic of China. 3. Research Center of Excellence in Regenerative Medicine, National Cheng Kung University, Tainan, Taiwan, Republic of China. 4. Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland. 5. Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland. 6. Viral Oncology Program, Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Abstract
Transmembrane proteins are responsible for many critical cellular functions and represent one of the largest families of drug targets. However, these proteins, especially multipass transmembrane proteins, are difficult to study because they must be embedded in a lipid bilayer to maintain their native conformations. The development of the virion display (VirD) technology enables transmembrane proteins to be integrated into the viral envelope of herpes simplex virus 1 (HSV-1). Combining high-throughput cloning, expression, and purification techniques, VirD technology has been applied to the largest set of human transmembrane proteins, namely G-protein-coupled receptors, and has allowed the identification of interactions that are both specific and functional. This article describes the procedures to integrate an open reading frame for any transmembrane protein into the HSV-1 genome and produce recombinant HSV-1 virus to ultimately generate pure VirD virions for biological and pharmaceutical studies.
Transmembrane proteins are responsible for many critical cellular functions and represent one of the largest families of drug targets. However, these proteins, especially multipass transmembrane proteins, are difficult to study because they must be embedded in a lipid bilayer to maintain their native conformations. The development of the virion display (VirD) technology enables transmembrane proteins to be integrated into the viral envelope of herpes simplex virus 1 (HSV-1). Combining high-throughput cloning, expression, and purification techniques, VirD technology has been applied to the largest set of human transmembrane proteins, namely G-protein-coupled receptors, and has allowed the identification of interactions that are both specific and functional. This article describes the procedures to integrate an open reading frame for any transmembrane protein into the HSV-1 genome and produce recombinant HSV-1 virus to ultimately generate pure VirD virions for biological and pharmaceutical studies.
Authors: Sang Ho Park; Sabrina Berkamp; Gabriel A Cook; Michelle K Chan; Hector Viadiu; Stanley J Opella Journal: Biochemistry Date: 2011-09-30 Impact factor: 3.162
Authors: Jens Frauenfeld; Robin Löving; Jean-Paul Armache; Andreas F-P Sonnen; Fatma Guettou; Per Moberg; Lin Zhu; Caroline Jegerschöld; Ali Flayhan; John A G Briggs; Henrik Garoff; Christian Löw; Yifan Cheng; Pär Nordlund Journal: Nat Methods Date: 2016-03-07 Impact factor: 28.547