Literature summary for 3.4.15.5 extracted from

Masuyer, G.; Cozier, G.E.; Kramer, G.J.; Bachmann, B.O.; Acharya, K.R.
Crystal structure of a peptidyl-dipeptidase K-26-DCP from Actinomycete in complex with its natural inhibitor (2016), FEBS J., 283, 4357-4369 .

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)	Astrosporangium hypotensionis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme free or in complex with natural microbial tripeptide product K-26, hanging drop vapour diffusion method, mixing of 10 mg/ml protein in 50 mM Tris, pH 8.0, and 150 mM NaCl, with reservoir solution consisting of 0.2 M MgCl2, 0.1 M sodium cacodylate, pH 6.5, and 20% PEG 8000, and equilibration over reservoir solution, at 16°C for 2 days. For the complex with K-26, the protein is preincubated with 5 mM K-26 at room temperature for 1 h before crystallisation. X-ray diffraction structure determination and analysis at 1.80-1.75 A resolution	Astrosporangium hypotensionis

Crystallization (Comment)

Organism

purified enzyme free or in complex with natural microbial tripeptide product K-26, hanging drop vapour diffusion method, mixing of 10 mg/ml protein in 50 mM Tris, pH 8.0, and 150 mM NaCl, with reservoir solution consisting of 0.2 M MgCl2, 0.1 M sodium cacodylate, pH 6.5, and 20% PEG 8000, and equilibration over reservoir solution, at 16°C for 2 days. For the complex with K-26, the protein is preincubated with 5 mM K-26 at room temperature for 1 h before crystallisation. X-ray diffraction structure determination and analysis at 1.80-1.75 A resolution

Astrosporangium hypotensionis

Inhibitors

Inhibitors	Comment	Organism	Structure
K-26 peptide	peptide K-26 from Astrosporangium hypotensionis is a good inhibitor of human angiotensin-I-converting enzyme (ACE), but a poor inhibitor of Astrosporangium hypotensionis K-26-DCP, binding analysis of K-26 peptide to K-26-DCP enzyme, overview. Ligand K-26 can bind to the active site of K-26-DCP. Unambiguous electron density is visible at the S1-S3 subsites within the substrate-binding channel of the enzyme in the complex crystal structure, structure modelling, overview. Comparison of the binding structure with human ACE	Astrosporangium hypotensionis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	dependent on, zinc dipeptidyl carboxypeptidase with a single catalytic zinc ion	Astrosporangium hypotensionis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
human angiotensin I + H2O	Astrosporangium hypotensionis	specific cleavage by enzyme K-26-DCP	?	-	?
human angiotensin I + H2O	Astrosporangium hypotensionis K-26	specific cleavage by enzyme K-26-DCP	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Astrosporangium hypotensionis	A0A1L1QK30	a soil-derived actinobacterium	-
Astrosporangium hypotensionis K-26	A0A1L1QK30	a soil-derived actinobacterium	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Astrosporangium hypotensionis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
human angiotensin I + H2O	specific cleavage by enzyme K-26-DCP	Astrosporangium hypotensionis	?	-	?
human angiotensin I + H2O	specific cleavage by enzyme K-26-DCP	Astrosporangium hypotensionis K-26	?	-	?

Subunits

Subunits	Comment	Organism
More	overall K-26-DCP is mostly a bundle of 25 alpha-helices accompanied by a five-stranded beta-sheet in subdomain II and a short double-stranded sheet in subdomain I	Astrosporangium hypotensionis

Synonyms

Synonyms	Comment	Organism
K-26-DCP	-	Astrosporangium hypotensionis
peptidyl-dipeptidase K-26-DCP	-	Astrosporangium hypotensionis

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.04	-	pH and temperature not specified in the publication	Astrosporangium hypotensionis	K-26 peptide

General Information

General Information	Comment	Organism
evolution	the peptidyl-dipeptidase K-26-DCP (K-26-DCP) of Astrosporangium hypotensionis is a zinc dipeptidyl carboxypeptidase (DCP) and an ancestral homologue of human angiotensin-I-converting enzyme (ACE). Enzyme K-26-DCP belongs to the M3 family of metalloproteases, that belong to the gluzincin class of enzymes (MEROPS database), which present a single catalytic zinc ion coordinated by the conserved HEXGH-binding motif. K-26-DCP shares some topological likeness with enzymes from the M2 family	Astrosporangium hypotensionis
metabolism	Astrosporangium hypotensionis has an unusual metabolism and is capable of producing complex peptide secondary metabolites. One of such peptides, K-26, presents a terminal phosphonic acid analogue of tyrosine, (R)-1-amino-2-(4-hydroxyphenyl)-ethylphosphonic acid ((R)-AHEP), and acts as good inhibitor of human angiotensin-I-converting enzyme (ACE), while K-26 is a poor inhibitor of bacterial DCP, including K-26-DCP produced by the same organism	Astrosporangium hypotensionis
additional information	the catalytic site is supported by subdomain I and is composed of the classical HEXXH Zn2+ coordination motif, made up by His 463, Glu 464, His 467, and completed by Glu 492. It resides at the centre of the protein deep within the central channel	Astrosporangium hypotensionis