𝔖 Bobbio Scriptorium
✦   LIBER   ✦

l-Pyroglutamyl-l-phenylalanyl-l-leucine-p-nitroanilide—A chromogenic substrate for thiol proteinase assay

✍ Scribed by I.Yu. Filippova; E.N. Lysogorskaya; E.S. Oksenoit; G.N. Rudenskaya; V.M. Stepanov

Book ID
102629389
Publisher
Elsevier Science
Year
1984
Tongue
English
Weight
354 KB
Volume
143
Category
Article
ISSN
0003-2697

No coin nor oath required. For personal study only.

✦ Synopsis

L-Pyroglutamyl-L-phenylalanyl-L-leucine-p-nitroanilide (PFLNA)--a convenient chromogenic substrate for assay of thiol proteinases papain, ficin, and bromelain--was prepared by enzymatic synthesis with chymotrypsin as a catalyst. The thiol proteinases hydrolyze PFLNA with the liberation of p-nitroaniline, estimated spectrophotometrically by its absorbance at 410 nm. The phenylalanine residue in the P2 position of PFLNA meets the specificity demands of thiol proteinases. The following values of Km were found for PFLNA hydrolysis: by papain, 0.34 mM; by ficin, 0.43 mM; by bromelain, 0.30 mM. This substrate was successfully applied to monitor thiol proteinase affinity chromatography on bacitracin-Sepharose, which resulted in a 2- to 4-fold purification from commercial preparations.

📜 SIMILAR VOLUMES

The hydrolysis of N-benzoyl-l-phenylalan
The hydrolysis of N-benzoyl-l-phenylalanyl-l-valyl-l-arginine-p-nitroanilide and its use as a substrate for the assay of cathepsin B
✍ John Butterworth; Judith J. Duncan 📂 Article 📅 1980 🏛 Elsevier Science 🌐 English ⚖ 428 KB

A calorimetric assay for the measurement of cathepsin B using N-benzoyl-r.-phenylalanyl-L-valyl+arginine-p-nitroanilide (S2160) as substrate has been developed. The technique is reproducible and simpler, quicker, safer, and more sensitive than that using cr-N-benzoyl-DL-arginine-P-naphthylamide as s