Literature summary for 3.4.21.75 extracted from

Pearce, K.; Overton, L.; Gampe, R.; Barrett, G.; Taylor, J.; McKee, D.; Campobasso, N.; Nolte, R.; Reid, R.
BacMam production and crystal structure of nonglycosylated apo human furin at 1.89 A resolution (2019), Acta Crystallogr. Sect. F, 75, 239-245 .

Search for more literature for 3.4.21.75

Application

Application	Comment	Organism
drug development	the deglycosylated active recombinant enzyme can be used for structure-guided drug discovery	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
gene FURIN, recombinant expression of glycosylated aa108-574-TEV-FLAG-His6-furin and nonglycosylated aa108-574-TEV-FLAG-His6 N387D/N440D furin mutant (ectodomains with a TEV cleavage site) from plasmid termed furin aa1-574 in CHO cells and in Spodoptera frugiperda Sf9 cells via BacMam virus transfection, typically producing a highly glycosylated, heterogeneous protein	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant furin mutant N387D/N440D in nonglycosylated apo-form, vapor diffusion method, 400 nl of 6 mg/ml of apo-furin (aa108-574-TEV-FLAG-His6 N387D/N440D) in 10 mM HEPES, pH 7.5, 150 mM NaCl, and 5 mM CaCl2, is mixed with 400 nl of crystallization solution containing 11-13% PEG 8000, 0.11-0.16 M potassium dihydrogen phosphate, and 0.1 M HEPES, pH 7.5, at 4°C, X-ray diffraction structure determination and analysis at 1.89 A resolution. The nonglycosylated furin protein reliably forms extremely durable apo crystals that diffract to high resolution, crystals are stable at 4°C for over one year. Digestion with TEV protease for the removal of the His6 and FLAG tags is not necessary for crystallization	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant furin mutant N387D/N440D in nonglycosylated apo-form, vapor diffusion method, 400 nl of 6 mg/ml of apo-furin (aa108-574-TEV-FLAG-His6 N387D/N440D) in 10 mM HEPES, pH 7.5, 150 mM NaCl, and 5 mM CaCl2, is mixed with 400 nl of crystallization solution containing 11-13% PEG 8000, 0.11-0.16 M potassium dihydrogen phosphate, and 0.1 M HEPES, pH 7.5, at 4°C, X-ray diffraction structure determination and analysis at 1.89 A resolution. The nonglycosylated furin protein reliably forms extremely durable apo crystals that diffract to high resolution, crystals are stable at 4°C for over one year. Digestion with TEV protease for the removal of the His6 and FLAG tags is not necessary for crystallization

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
additional information	expression and purification of nonglycosylated human furin using the BacMam technology and site-directed mutagenesis of the glycosylation sites. Recombinant nonglycosylated furin produced using this system retains full proteolytic activity indistinguishable from that of the glycosylated protein	Homo sapiens
N387D/N440D	site-directed mutagenesis, mutation of glycosylation sites	Homo sapiens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ca2+	dependent on	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P09958	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	-	Homo sapiens

Purification (Commentary)

Purification (Comment)	Organism
recombinant FLAG/His6-tagged furin ectodomain from CHO cells or Spodoptera frugiperda Sf9 cells by nickel affinity chromatography, ultrafiltration, and gel filtration, to homogeneity	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
brain	-	Homo sapiens	-
carcinoma cell	-	Homo sapiens	-
lung	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5-carboxyfluorescein-Gln-Arg-Val-Arg-Arg-Ala-Val-Gly-Ile-Asp-Lys(5-carboxytetramethylrhodamine)-OH + H2O	-	Homo sapiens	?	-	?

Subunits

Subunits	Comment	Organism
homodimer	2 * 53446, recombinant prodomain-processed, nonglycosylated aa108-574-TEV-FLAG-His6 N387D/N440D mutant, mass spectrometry	Homo sapiens

Synonyms

Synonyms	Comment	Organism
PCSK3	-	Homo sapiens
proprotein convertase subtilisin/kexin 3	-	Homo sapiens

General Information

General Information	Comment	Organism
evolution	furin belongs to the proprotein convertase subtilisin/kexin (PCSK) family of enzymes play a crucial role in processing and trafficking of a wide variety of precursor proteins such as hormones, receptors, and enzymes	Homo sapiens
malfunction	dysregulation of furin is implicated in numerous disease states, including cancer and fibrosis	Homo sapiens
physiological function	furin is a calcium-dependent serine endoprotease that processes a wide variety of proproteins involved in cell function and homeostasis	Homo sapiens