Furin Structures

Overview to structures

(Pearce Reid 2019 pmid: 30950824) - truncated, heterogeneous glycosylated mouse furin (ca. 60 kDa) produced in Chinese hamster ovary (CHO) cells was used to determine the first crystal structure of furin in complex with the decanoyl-Arg-Val-Lys-Arg-chloro- methyl ketone inhibitor at 2.6 A resolution (Henrich et al., 2003).

Human furin with two different inhibitors were reported at 2.3 and 2.7 Aesolution (Dahms et al., 2014). In this case, uniformly glycosylated, soluble furin protein was prepared from an engineered human embryonic kidney cell line (HEK293S Gnt-),

recently reported higher resolution structures of glycosylated human furin at 1.8 to 2.0 A solution in unliganded and ligand-bound functional forms (Dahms et al., 2016, 2018). - attempts at preparing reliable diffraction-quality crystals of glycosylated human furin were unsuccessful.

homogeneous protein containing aspartic acid substitutions at two known N-linked glycosylation sites, Asn387 and Asn440 (N387D/N440D), used for biochemical and high-throughput crystallographic studies (Ames et al., 2007; Scott et al., 2007; Dukkipati et al., 2008). This nonglycosylated form of the furin ecto-domain has similar enzymatic properties to the native protein and readily crystallizes in the apo form, providing an excellent system for small-molecule soaks.

Structural Dynamics

(Dahms 2016 PNAS pmid: 27647913) Furin resolved in 4 different structures (- /+ Ca); (- /+ Substrates) - provide hints on how to do docking

Furin cristal structures with and without inhibitors

For PharmAI: Could you 1. select Furin structures to look whether Camostat and Nafamostat do also inhibit Furin (TMPRSS2 - S protein processing); 2. Screen the FDA approved small molecules for Furin binding; 3. Screen a large small molecule library

(Dahms Than 2014 pmid: 24666235) - Peptidomimetic compounds based on a phenylacetyl-ArgVal-Arg-4-(amidomethyl)benzamidine (Phac-RVR-4-Amba) core structure 15 belong to the strongest noncovalent inhibitors available so far. (PDB entry 4omd )

(Hardes Steinmetzer 2015 pmid: 25974265) structure with peptidomimetic inhibitor. inhibition assay with influenza.

(Dahms Than 2016 pmid: 27647913) - furin in different functional states were determined to 1.8–2.0 Å. These, together with biochemical data and modeling by molecular dynamics calculations, suggest essential elements underlying its unusually high substrate specificity. Furin shows a complex activation mechanism and exists in at least four defined states: (i) the “off state,” incompatible with substrate binding as seen in the unliganded enzyme; (ii) the active “on state” seen in inhibitor-bound furin; and the respective (iii) calcium-free and (iv) calcium-bound forms

(Dahms Than 2016 SciRep pmid: 27670069) - structure with an inhibitory camelid antibody

(Dahms Than 2017 pmid: 28402100)

(Lam Steinmetzer 2019 pmid: 30680958) - structure with a hexapeptide - 6EQX

(Pearce Reid 2019 pmid: 30950824) - 1.89A structure BacMam construct. 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. I will receive the BacMam construct from GSK 3-20 apo furin structure 4z2a; liganded furin (PDB entry 4omd Structure of Than 2014). Apo furin shows differing rotamers for His154 and Ser368.

Than and colleagues made 19 Furin structures / 10 structures <2A /