Literature summary for 3.4.24.13 extracted from

Chi, Y.C.; Rahkola, J.T.; Kendrick, A.A.; Holliday, M.J.; Paukovich, N.; Roberts, T.S.; Janoff, E.N.; Eisenmesser, E.Z.
Streptococcus pneumoniae IgA1 protease a metalloprotease that can catalyze in a split manner in vitro (2017), Protein Sci., 26, 600-610 .

Search for more literature for 3.4.24.13

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminally tagged wild-type and mutant active mature enzymes in Escherichia coli strain BL21(DE3). The constructs contain an N-terminal 6xHis tag and a C-terminal MBP tag harboring an N-terminal TEV cleavage site and a C-terminal thrombin cleavage site, respectively, or the constructs contain a C-terminal His6-tag with thrombin cleavage site	Streptococcus pneumoniae

Protein Variants

Protein Variants	Comment	Organism
E1605A	site-directed mutagenesis, the IgA1P mutation completely abrogates the proteolytic activity	Streptococcus pneumoniae
E1628A	site-directed mutagenesis, the IgA1P mutation reduces the proteolytic activity of the enzyme	Streptococcus pneumoniae
E1661A	site-directed mutagenesis, the IgA1P mutation shows proteolytic activity similar to the wild-type	Streptococcus pneumoniae
E1695A	site-directed mutagenesis, the IgA1P mutation shows proteolytic activity similar to the wild-type	Streptococcus pneumoniae
E1698A	site-directed mutagenesis, the IgA1P mutation shows proteolytic activity similar to the wild-type	Streptococcus pneumoniae
additional information	proteolytic preparation of wild-type and mutant N- and C-terminal constructs of the recombinant enzyme: IgA1P154-1963, E1605A-IgA1P, IgA1P154-1611, IgA1P1612-1963, and IgA1P1593-1963 and the combination of IgA1P154-1611 and IgA1P1612-196. The Streptococcus pneumoniae IgA1P activity can be reconstituted in trans by the independently purified N-terminal and C-terminal domains	Streptococcus pneumoniae

Inhibitors

Inhibitors	Comment	Organism	Structure
EDTA	-	Streptococcus pneumoniae

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cell surface	-	Streptococcus pneumoniae	9986	-
extracellular	-	Streptococcus pneumoniae	-	-
additional information	the mature Streptococcus pneumoniae IgA1P form is both attached to the bacterial cell surface and released in its full form	Streptococcus pneumoniae	-	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	dependent on, metalloproteinase, the enzyme owns a conserved HExxH Zn-binding motif (1604-1608), which coordinates Zn2+ via residues His1604 and His1608, and a third one, and provides the catalytic base, Glu1605	Streptococcus pneumoniae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
human immunoglobulin A1 + H2O	Streptococcus pneumoniae	specific cleavage in the hinge region	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Streptococcus pneumoniae	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
proteolytic modification	the fully mature form of Streptococcus pneumoniae IgA1P comprises residues 154-1963	Streptococcus pneumoniae

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally tagged wild-type and mutant active mature enzymes from Escherichia coli strain BL21(DE3) by nickel and amylose affinity chromatography, MBP-tag cleavage through TEV and the His-tag through thrombin, and gel filtration. Recombinant C-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography an, tag cleavage through thrombin, and gel filtration	Streptococcus pneumoniae

Purification (Comment)

Organism

recombinant N-terminally tagged wild-type and mutant active mature enzymes from Escherichia coli strain BL21(DE3) by nickel and amylose affinity chromatography, MBP-tag cleavage through TEV and the His-tag through thrombin, and gel filtration. Recombinant C-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography an, tag cleavage through thrombin, and gel filtration

Streptococcus pneumoniae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
human immunoglobulin A1 + H2O	specific cleavage in the hinge region	Streptococcus pneumoniae	?	-	?
human immunoglobulin A1 + H2O	specific cleavage in the hinge region, the C-terminal region of enzyme IgA1P is necessary for cleavage of IgA1	Streptococcus pneumoniae	?	-	?

Synonyms

Synonyms	Comment	Organism
IgA1 protease	-	Streptococcus pneumoniae
IgA1P	-	Streptococcus pneumoniae
Streptococcus pneumoniae IgA1 protease	-	Streptococcus pneumoniae

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at room temperature	Streptococcus pneumoniae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Streptococcus pneumoniae

General Information

General Information	Comment	Organism
additional information	the conserved HExxH motif (amino acid residues 1604-1608) coordinates Zn2+ and provides the catalytic base, Glu1605. Residue Glu1628 also contributes to IgA1P catalytic activity. The C-terminal domain, IgA1P1612-1963, exhibits no detectable binding to IgA1. The Streptococcus pneumoniae IgA1P N- and C-terminal regions are catalytically active when combined in trans	Streptococcus pneumoniae
physiological function	IgA1 protease (IgA1P) specifically cleaves human immunoglobulin A1 (IgA1) at the hinge region dissociating the effector Fc region while the IgA1 FAB remains bound to the bacterial surface. This cleavage promotes infection by two mechanisms. Firstly, Fc receptors on neutrophils can no longer recognize the cleaved and released IgA1 Fc to bind and kill the bacterial cells. Secondly, the IgA1 FAB fragment remaining on the Streptococcus pneumoniae cell surface enhances Streptococcus pneumoniae binding to epithelial cells and can no longer prevent colonization	Streptococcus pneumoniae