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Literature summary for 3.4.24.16 extracted from

  • Hines, C.S.; Ray, K.; Schmidt, J.J.; Xiong, F.; Feenstra, R.W.; Pras-Raves, M.; de Moes, J.P.; Lange, J.H.; Melikishvili, M.; Fried, M.G.; Mortenson, P.; Charlton, M.; Patel, Y.; Courtney, S.M.; Kruse, C.G.; Rodgers, D.W.
    Allosteric inhibition of the neuropeptidase neurolysin (2014), J. Biol. Chem., 289, 35605-35619.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.24.16

Cloned(Commentary)

Cloned (Comment) Organism
recombinant enzyme expression in Escherichia coli Rattus norvegicus

Crystallization (Commentary)

Crystallization (Comment) Organism
purified enzyme with bound inhibitor R2, hanging drop vapor diffusion, mixing 2 ml of 48 mM protein in the presence of 5fold stoichiometric excess of inhibitor R2 with 1 ml of well solution containing 0.1 M HEPES, pH 7.0, 0.1 M LiSO4,2 mM 2-mercaptoethanol, and 12-15% PEG 4000, 4°C, X-ray diffraction structure determination and analysis at 2.8 A resolution Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
E475A site-directed mutagenesis, mutation of the catalytic glutamate residue, a catalytically compromised enzyme mutant that sediments more rapidly in the presence of saturating amounts of the dynorphin A(1-8) substrate compared to the wild-type enzyme Rattus norvegicus
H160A site-directed mutagenesis, mutation of a surface histidine involved in a divalent cation-mediated lattice contact in the original neurolysin crystals Rattus norvegicus

Inhibitors

Inhibitors Comment Organism Structure
1-adamantan-2-yl-3-[2-[2,3-bis-(2-chloro-phenyl)-pyrazolidin-1-yl]-2-oxo-ethyl]-urea racemic Rattus norvegicus
3-[(2S)-1-[(3R)-3-(2-chlorophenyl)-2-(2-fluorophenyl)pyrazolidin-1-yl]-1-oxopropan-2-yl]-1-(adamantan-2-yl)urea R-stereomer, allosteric inhibition of neurolysin, mixed-type inhibition kinetics, the bound inhibitor disrupts activity by preventing a hinge-like motion associated with substrate binding and catalysis. The inhibitor reverses a substrate-associated conformational change Rattus norvegicus
additional information inhibitor synthesis and evaluation, different kinetic inhibition models, binding structures, modeling, overview Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus P42676
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-aminobenzoyl-dynorphin A(1-8)-N-(2,4-dinitrophenyl)-ethylenediamine + H2O binding structure and conformational effects on the enzyme compared to inhibitor R2 Rattus norvegicus ?
-
 ?
2-aminobenzoyl-neurotensin-N-(2,4-dinitrophenyl)-ethylenediamine + H2O binding structure and conformational effects on the enzyme compared to inhibitor R2 Rattus norvegicus ?
-
 ?
additional information models of substrate binding in the neurolysin active site channel, overview Rattus norvegicus ?
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Rattus norvegicus

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.0066
-
 pH 7.5, 37°C Rattus norvegicus 1-adamantan-2-yl-3-[2-[2,3-bis-(2-chloro-phenyl)-pyrazolidin-1-yl]-2-oxo-ethyl]-urea
0.0075
-
 pH 7.5, 37°C Rattus norvegicus 3-[(2S)-1-[(3R)-3-(2-chlorophenyl)-2-(2-fluorophenyl)pyrazolidin-1-yl]-1-oxopropan-2-yl]-1-(adamantan-2-yl)urea

General Information

General Information Comment Organism
physiological function neuropeptidases specialize in the hydrolysis of the small bioactive peptides that play a variety of signaling roles in the nervous and endocrine systems. Neuropeptidase neurolysin helps control the levels of the dopaminergic circuit modulator neurotensin and is a member of a fold group that includes the antihypertensive target angiotensin converting enzyme Rattus norvegicus