Download MendeleyBacMam production and crystal structure of nonglycosylated apo human furin at 1.89 angstrom resolution.
Pearce, K.H., Overton, L.K., Gampe, R.T., Barrett, G.B., Taylor, J.D., McKee, D.D., Campobasso, N., Nolte, R.T., Reid, R.A.(2019) Acta Crystallogr F Struct Biol Commun 75: 239-245
- PubMed: 30950824
- DOI: https://doi.org/10.1107/S2053230X19001419
- Primary Citation of Related Structures:
4Z2A - PubMed Abstract:
Furin, also called proprotein convertase subtilisin/kexin 3 (PCSK3), is a calcium-dependent serine endoprotease that processes a wide variety of proproteins involved in cell function and homeostasis. Dysregulation of furin has been implicated in numerous disease states, including cancer and fibrosis. Mammalian cell expression of the furin ectodomain typically produces a highly glycosylated, heterogeneous protein, which can make crystallographic studies difficult. Here, the expression and purification of nonglycosylated human furin using the BacMam technology and site-directed mutagenesis of the glycosylation sites is reported. Nonglycosylated furin produced using this system retains full proteolytic activity indistinguishable from that of the glycosylated protein. Importantly, the nonglycosylated furin protein reliably forms extremely durable apo crystals that diffract to high resolution. These crystals can be soaked with a wide variety of inhibitors to enable a structure-guided drug-discovery campaign.
- Eshelman School of Pharmacy, Division of Chemical Biology and Medicinal Chemistry, Center for Integrative Chemical Biology and Drug Discovery, University of North Carolina at Chapel Hill, 125 Mason Farm Road, Chapel Hill, NC 27599, USA.
Organizational Affiliation:
