Literature summary for 3.4.21.112 extracted from

Da Palma, J.; Burri, D.; Oppliger, J.; Salamina, M.; Cendron, L.; De Laureto, P.; Seidah, N.; Kunz, S.; Pasquato, A.
Zymogen activation and subcellular activity of subtilisin kexin isozyme 1/site 1 protease (2014), J. Biol. Chem., 289, 35743-35756.

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Cloned(Commentary)

Cloned (Comment)	Organism
transient expression of C-terminally His6-tagged full-length wild-type enzyme and a soluble enzyme variant truncated before the transmembrane domain, the catalytically inactive mutant H249A, B'/B, B' mutants of enzyme in HEK-293T cells, secretion of proteins. Wild-type and the DELTA AC SKI-1/S1P FL mutant are expressed in the SKI-I/S1P-deficient CHO cell line SRD12B, which lacks active endogenous enzyme	Homo sapiens

Cloned (Comment)

Organism

transient expression of C-terminally His6-tagged full-length wild-type enzyme and a soluble enzyme variant truncated before the transmembrane domain, the catalytically inactive mutant H249A, B'/B, B' mutants of enzyme in HEK-293T cells, secretion of proteins. Wild-type and the DELTA AC SKI-1/S1P FL mutant are expressed in the SKI-I/S1P-deficient CHO cell line SRD12B, which lacks active endogenous enzyme

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
H249A	site-directed mutagenesis, catalytically inactive mutant	Homo sapiens
additional information	the soluble form of the enzyme is truncated before the transmembrane domain and comprised the ectodomain, followed by a C-terminal V5 tag, i.e. DELTA AC SKI-1/S1P BTMD. Extended mutagenesis performed on a region proximal to the C site results in normal SKI-1/S1P maturation at the B'/B intermediate state, suggesting that processing at B'/B either precedes or occurs independently of C site cleavage. The combined mutation at the C and C' sites results in marked reduction of the mature C form indicating that, similar to B'/B, mutation of both C and C' processing sites is required to prevent maturation	Homo sapiens
R130E/R134E	site-directed mutagenesis, the double mutations at the B'/B site prevents autoprocessing	Homo sapiens
R134E	site-directed mutagenesis, the single prodomain mutant, at the B autoprocessing site, allows maturation comparably with the wild-type enzyme	Homo sapiens
R160E	site-directed mutagenesis	Homo sapiens
R163E/R164E	site-directed mutagenesis	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
Golgi membrane	-	Homo sapiens	139	-
membrane	transmembrane protein	Homo sapiens	16020	-
additional information	the truncated mutant DELTA AC SKI-1/S1P BTMD, truncated before the transmembrane domain, stays in the endoplasmic reticulum	Homo sapiens	-	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Homo sapiens	zymogen activation of the enzyme involving sequential autocatalytic processing of its N-terminal prodomain at sites B'/B followed by the C'/C sites. Enzyme autoprocessing results in intermediates whose catalytic domain remains associated with prodomain fragments of different lengths. All incompletely matured intermediates of SKI-1/S1P show full catalytic activity toward cellular substrates, whereas optimal cleavage of viral glycoproteins depends on B'/B processing. Incompletely matured forms of SKI-1/S1P further process cellular and viral substrates in distinct subcellular compartments	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	Q14703	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	-	Homo sapiens
proteolytic modification	zymogen activation of the enzyme involves sequential autocatalytic processing of its N-terminal prodomain at sites B'/B followed by the C'/C sites. Autocatalytic maturation by sequential cleavages of the N-terminal pro-domain first at sites B'/B (RKVF2RSLK1372), followed by site C (RRLL1862), with crucial roles for R and V/L residues at P4 and P2 (fourth and second residue upstream the scissile bond, respectively). Enzyme autoprocessing results in intermediates whose catalytic domain remains associated with prodomain fragments of different lengths. 9 Amino acid residues at the cleavage site (P1-P8) and P1' are necessary and sufficient to define the subcellular location of processing and to determine to what extent processing of a substrate depends on SKI-1/S1P maturation. Mature enzyme retains prodomain fragments of different lengths, overview. The post-translational modifications are crucial for the subcellular localization of autoprocessing intermediates of the enzyme	Homo sapiens

Purification (Commentary)

Purification (Comment)	Organism
recombinant C-terminally His6-tagged wild-type mutant enzyme from HEK-293T cell medium by immobilized metal affinity chromatography	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	zymogen activation of the enzyme involving sequential autocatalytic processing of its N-terminal prodomain at sites B'/B followed by the C'/C sites. Enzyme autoprocessing results in intermediates whose catalytic domain remains associated with prodomain fragments of different lengths. All incompletely matured intermediates of SKI-1/S1P show full catalytic activity toward cellular substrates, whereas optimal cleavage of viral glycoproteins depends on B'/B processing. Incompletely matured forms of SKI-1/S1P further process cellular and viral substrates in distinct subcellular compartments	Homo sapiens	?	-	?
additional information	substrate specificity and activity of wild-type enzyme and pro-domain mutants towards cellular and viral substrates, construction and evaluation of a cell-based chimeric protein molecular sensor, containing the LASVGPC cleavage site IYISRRLL-/-G, to monitor endogenous enzyme activity, overview	Homo sapiens	?	-	?

Synonyms

Synonyms	Comment	Organism
SKI-1/S1P	-	Homo sapiens
subtilisin kexin isozyme 1/site 1 protease	-	Homo sapiens

General Information

General Information	Comment	Organism
additional information	9 amino acid residues at the cleavage site (P1-P8) and P1' are necessary and sufficient to define the subcellular location of processing and to determine to what extent processing of a substrate depends on SKI-1/S1P maturation	Homo sapiens
physiological function	the enzyme plays crucial roles in cellular homeostatic functions and is hijacked by pathogenic viruses for the processing of their envelope glycoproteins. Zymogen activation of the enzyme involves sequential autocatalytic processing of its N-terminal prodomain at sites B'/B followed by the C'/C sites. All incompletely matured intermediates of SKI-1/S1P showed full catalytic activity toward cellular substrates, whereas optimal cleavage of viral glycoproteins depended on B'/B processing. Incompletely matured forms of SKI-1/S1P further process cellular and viral substrates in distinct subcellular compartments	Homo sapiens