Literature summary for 3.4.21.76 extracted from

Kuhl, A.; Korkmaz, B.; Utecht, B.; Kniepert, A.; Schoenermarck, U.; Specks, U.; Jenne, D.E.
Mapping of conformational epitopes on human proteinase 3, the autoantigen of Wegener's granulomatosis (2010), J. Immunol., 185, 387-399.

Search for more literature for 3.4.21.76

Application

Application	Comment	Organism
medicine	neutrophil-activating potential of cANCAs can vary in patients, depending on the association rates, target specificities, and titers of antibodies, as well as on local concentrations of human PR3 inhibitors	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
full-length cDNA amplified, enterokinase cleavage site (DDDDK) introduced at the N terminus of PR3. PCR product digested with PmlI/AgeI and cloned into the reading frame of the S-tag, resulting in construct pcDNA5/FRT/hPR3-H6. Human/gibbon chimeras performed by digestion of pcDNA5/FRT/PR3-H6 with BlpI and AgeI and exchange of the respective cDNA segments. Expression in Flp-in HEK293 cells	Homo sapiens
granulocyte cDNA amplified, PCR fragment reamplified and cloned into pcDNA5/FRT. Human/gibbon chimeras performed by digestion of pcDNA5/FRT/PR3-H6 with BlpI and AgeI and exchange of the respective cDNA segments. Chemically synthesized 390-bp human PR3 fragment digested with PmlI/BlpI and cloned into the same sites of the pcDNA5/FRT/gPR3-H6 plasmid to reconstruct the human CLB12.8 epitope in gibbon PR3 (gibbon proteinase 3 variant 1). A further modification of this variant by replacing the A116 to Q119 tetrapeptide with the respective opossum residues Q-V-A-S. To this end, a DNA fragment chemically synthesized, digested with BlpI and BstEII, and subcloned into the same sites of the gPR3v1 plasmid. The two glycosylation sites Asn113 and Asn159 mutated to lysine residues in the gibbon proteinase 3 variant 2. Expression in Flp-in HEK293 cells	Hylobates pileatus

Cloned (Comment)

Organism

full-length cDNA amplified, enterokinase cleavage site (DDDDK) introduced at the N terminus of PR3. PCR product digested with PmlI/AgeI and cloned into the reading frame of the S-tag, resulting in construct pcDNA5/FRT/hPR3-H6. Human/gibbon chimeras performed by digestion of pcDNA5/FRT/PR3-H6 with BlpI and AgeI and exchange of the respective cDNA segments. Expression in Flp-in HEK293 cells

Homo sapiens

granulocyte cDNA amplified, PCR fragment reamplified and cloned into pcDNA5/FRT. Human/gibbon chimeras performed by digestion of pcDNA5/FRT/PR3-H6 with BlpI and AgeI and exchange of the respective cDNA segments. Chemically synthesized 390-bp human PR3 fragment digested with PmlI/BlpI and cloned into the same sites of the pcDNA5/FRT/gPR3-H6 plasmid to reconstruct the human CLB12.8 epitope in gibbon PR3 (gibbon proteinase 3 variant 1). A further modification of this variant by replacing the A116 to Q119 tetrapeptide with the respective opossum residues Q-V-A-S. To this end, a DNA fragment chemically synthesized, digested with BlpI and BstEII, and subcloned into the same sites of the gPR3v1 plasmid. The two glycosylation sites Asn113 and Asn159 mutated to lysine residues in the gibbon proteinase 3 variant 2. Expression in Flp-in HEK293 cells

Hylobates pileatus

Protein Variants

Protein Variants	Comment	Organism
V35M/S38AN/I38BP/Q74R/N113K/N159K/119-122QVAS	results in the gibbon proteinase 3 variant 2. Is no longer able to interact with the mouse antibodies 4A5 and WGM2, but retains its binding to CLB12.8	Hylobates pileatus
V35M/S38AN/L39BP/Q74R	results in the gibbon proteinase 3 variant 1. The binding site for mouse antibody CLB12.8, but not that for MCPR3-2, is reconstituted. Binds also to mouse antibody 6A6. Like MCPR3-2, WGM3 binds neither to gibbon PR3 nor to the humanized gibbon proteinase 3 variant 1	Hylobates pileatus

Inhibitors

Inhibitors	Comment	Organism
alpha1-protease inhibitor	-	Homo sapiens
alpha1-protease inhibitor	purified human alpha1-protease inhibitor inhibits gibbon PR3 and forms a covalently linked complex with it	Hylobates pileatus
elafin	is inhibited by purified human elafin, a canonical low molecular weight inhibitor of human PR3	Hylobates pileatus

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-
Hylobates pileatus	-	-	-
Macaca mulatta	-	-	-
Pan troglodytes verus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
6 x His-tagged proPR3 purified using affinity chromatography	Homo sapiens
6 x His-tagged proPR3 purified using affinity chromatography	Hylobates pileatus

Source Tissue

Source Tissue	Comment	Organism	Textmining
blood	-	Macaca mulatta	-
blood	-	Hylobates pileatus	-
blood	-	Pan troglodytes verus	-
neutrophil	-	Macaca mulatta	-
neutrophil	-	Hylobates pileatus	-
neutrophil	-	Pan troglodytes verus	-
serum	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
Abz-VADCADQ-(2,4-dinitrophenyl)ethylenediamine + H2O	-	Hylobates pileatus	?	-	?
Abz-VADCADQ-N-(2,4-dinitrophenyl)ethylenediamine + H2O	-	Homo sapiens	?	-	?
Boc-Ala-Pro-Nva-4-chloro-thiobenzyl ester + H2O	-	Homo sapiens	?	-	?
Boc-Ala-Pro-Nva-4-chloro-thiobenzyl ester + H2O	-	Hylobates pileatus	?	-	?

Synonyms

Synonyms	Comment	Organism
PR3	-	Homo sapiens
PR3	-	Macaca mulatta
PR3	-	Hylobates pileatus
PR3	-	Pan troglodytes verus
proteinase 3	-	Homo sapiens
proteinase 3	-	Macaca mulatta
proteinase 3	-	Hylobates pileatus
proteinase 3	-	Pan troglodytes verus

General Information

General Information	Comment	Organism
physiological function	almost complete absence of anti-neutrophil cytoplasmic antibody (cANCA) binding to rhesus monkey PR3	Pan troglodytes verus
physiological function	anti-neutrophil cytoplasmic antibodies (cANCAs) from Wegeners granulomatosis patients at least in part recognize similar surface structures as do mouse antibodies and compete with the binding of alpha1-protease inhibitor to PR3	Homo sapiens
physiological function	gibbon PR3 shows an intermediate anti-neutrophil cytoplasmic antibody (cANCA) binding pattern, whereas some anti-PR3 mouse antibodies and some patient sera recognize the antigen. The cANCAs from Wegeners granulomatosis patients at least in part recognize similar surface structures as do mouse antibodies and compete with the binding of alpha1-protease inhibitor to PR3	Hylobates pileatus
physiological function	macaque PR3 shows nearly no reactivity to anti-neutrophil cytoplasmic antibodies (cANCAs)	Macaca mulatta