- PDB-1zg7: Crystal Structure of 2-(5-{[amino(imino)methyl]amino}-2-chlorophe... -
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Entry
Database: PDB / ID: 1zg7
Title
Crystal Structure of 2-(5-{[amino(imino)methyl]amino}-2-chlorophenyl)-3-sulfanylpropanoic acid Bound to Activated Porcine Pancreatic Carboxypeptidase B
Components
procarboxypeptidase B
Keywords
HYDROLASE / CARBOXYPEPTIDASE B / EXOPEPTIDASE / thiol based inhibitor
Function / homology
Function and homology information
carboxypeptidase B / metallocarboxypeptidase activity / cytoplasmic vesicle / proteolysis / extracellular space / zinc ion binding Similarity search - Function
HETEROGEN THE INHIBITION CONSTANT (KI APPARENT) OF 2-(5-{[AMINO (IMINO)METHYL] AMINO}-2- ...HETEROGEN THE INHIBITION CONSTANT (KI APPARENT) OF 2-(5-{[AMINO (IMINO)METHYL] AMINO}-2-CHLOROPHENYL)-3-SULFANYLPROPANOIC ACID WAS MEASURED AS 7 NM AGAINST PORCINE PANCREATIC CARBOXYPEPTIDASE B. THE ASSAY FOR INHIBITION OF PORCINE PANCREATIC CPB (SIGMA, ST. LOUIS, MO) WAS PERFORMED IN A 96 OR 384-WELL FORMAT ADAPTED FROM PUBLISHED PROTOCOLS. PURIFIED PP-CPB (2 NM) WAS INCUBATED WITH TEST COMPOUNDS IN 20 MM HEPES PH 7.4, 150 MM NACL, 5 MM CACL2 FOR 2 MINUTES PRIOR TO THE ADDITION OF THE 0.6 MM HIPPURYL-L -ARGININE SUBSTRATE. AFTER 30 MINUTES AT ROOM TEMPERATURE, THE AMOUNT OF SUBSTRATE HYDROLYZED WAS DETERMINED BY CONVERSION OF THE PRODUCT, HIPPURIC ACID, TO A CHROMOGEN WITH SODIUM PHOSPHATE BUFFER, PH 8.3 (F.C. 0.8 MM) AND CYANURIC CHLORIDE/DIOXANE (F.C. 0.9% W/V) UNDER CHEMICAL FUME HOOD. FOLLOWING CENTRIFUGATION OF THE MICROTITER PLATES AND TRANSFER OF THE SUPERNATANT TO A CLEAN PLATE, ABSORBANCE OF THE SUPERNATANT IS READ AT 382 NM. THE IC50 OF THE COMPOUND WAS DETERMINED USING THE 4-PARAMETER EQUATION FROM AN 8-POINT DOSE RESPONSE CURVE, EACH COMPOUND TESTED IN DUPLICATE.
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