MilliporeSigma
  • Home
  • Search Results
  • Evaluation of cathepsin B activity for degrading collagen IV using a surface plasmon resonance method and circular dichroism spectroscopy.

Evaluation of cathepsin B activity for degrading collagen IV using a surface plasmon resonance method and circular dichroism spectroscopy.

Journal of pharmaceutical and biomedical analysis (2014-03-19)
Atsushi Shoji, Mitsutaka Kabeya, Yuuki Ishida, Akio Yanagida, Yoichi Shibusawa, Masao Sugawara
ABSTRACT

Evaluation of cathepsin B activities for degrading collagen IV and heat-denatured collagen IV (gelatin) were performed by surface plasmon resonance (SPR) and circular dichroism (CD) measurements. The optimal pH of cathepsin B activity for degrading each substrate was around 4.0. The ΔRU(15 min), which is a decrease in the SPR signal at 15 min after injection of cathepsin B, was smaller for collagen IV than for heat-denatured collagen IV owing to the presence of triple-helical conformation. An unstable nature of the triple-helical conformation of collagen IV at pH 4.0 was shown by the CD study. Degrading collagen IV by cathepsin B was facilitated owing to a local unwinding of the triple-helical conformation caused by proteolytic cleavage of the non-helical region. The concentration dependence of the initial velocity for degrading collagen IV by cathepsin B at pH 4.0 was biphasic, showing that cathepsin B at low concentration exhibits exopeptidase activity, while the enzyme at high concentration exhibits endopeptidase activity. The kinetic parameters for the exopeptidase activity of cathepsin B toward collagen IV and heat-treated collagen IV were evaluated and discussed in terms of the protease mechanism.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Boric acid, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
Sodium acetate trihydrate, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
Citric acid, BioUltra, anhydrous, ≥99.5% (T)
Sigma-Aldrich
Citric acid, ≥99.5%, FCC, FG
Sigma-Aldrich
2-Hydroxybutyric acid sodium salt, 97%
Sigma-Aldrich
Citric acid, ACS reagent, ≥99.5%
Sigma-Aldrich
N-Hydroxysuccinimide, purum, ≥97.0% (T)
Sigma-Aldrich
Boric acid, 99.999% trace metals basis
Sigma-Aldrich
Boric acid, 99.97% trace metals basis
Sigma-Aldrich
N-Hydroxysuccinimide, 98%
Sigma-Aldrich
Sodium acetate trihydrate, meets USP testing specifications
Sigma-Aldrich
Sodium acetate trihydrate, BioXtra, ≥99.0%
Sigma-Aldrich
3-(2-Pyridyldithio)propionic acid N-hydroxysuccinimide ester, ≥95%, powder
Sigma-Aldrich
Citric acid, anhydrous, suitable for cell culture, suitable for plant cell culture
Sigma-Aldrich
Boric acid, BioXtra, ≥99.5%
Sigma-Aldrich
Citric acid, 99%
Sigma-Aldrich
Boric acid, suitable for electrophoresis, ≥99.5%
Sigma-Aldrich
Boric acid, tablet, 1 g boric acid per tablet
Sigma-Aldrich
Boric acid, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
Supelco
Citric acid, Anhydrous, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Tris(2-carboxyethyl)phosphine hydrochloride solution, 0.5 M, pH 7.0(aqueous solution; pH was adjusted with ammonium hydroxide)
USP
Citric acid, United States Pharmacopeia (USP) Reference Standard
Supelco
Benalaxyl-M, PESTANAL®, analytical standard
Supelco
Citric acid, certified reference material, TraceCERT®
Sigma-Aldrich
Boric acid, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., buffer substance, ≥99.8%
Sigma-Aldrich
Sodium acetate trihydrate, ACS reagent, ≥99%
Sigma-Aldrich
Sodium acetate trihydrate, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, E262, 99.0-101.0% (calc. to the dried substance), ≤0.00002% Al
Sigma-Aldrich
Citric acid, meets analytical specification of Ph. Eur., BP, USP, E330, anhydrous, 99.5-100.5% (based on anhydrous substance)
Sigma-Aldrich
Boric acid, ReagentPlus®, ≥99.5%
Citric acid, anhydrous, European Pharmacopoeia (EP) Reference Standard