PMID- 22542927
OWN - NLM
STAT- MEDLINE
DCOM- 20121025
LR  - 20131121
IS  - 1873-4235 (Electronic)
IS  - 0956-5663 (Linking)
VI  - 36
IP  - 1
DP  - 2012 Jun-Jul
TI  - Monitoring of proteolytic enzyme activity using phase transition-based peptide 
      arrays.
PG  - 147-53
LID - 10.1016/j.bios.2012.04.004 [doi]
AB  - We have developed an assay using peptide arrays based on phase transition from 
      the glass substrate to the liquid for monitoring quantitative protease activity 
      in real-time. Peptide arrays were fabricated using a bifunctional cross-linker, 
      N-[gamma-maleimidobutyryloxy] sulfosuccinimide ester, and a substrate peptide 
      containing two functional groups, cysteine and tetramethyl-6-carboxyrhodamine 
      (TAMRA) on the C- and N-terminus, respectively. The phase transition-based 
      peptide arrays were characterized by analyzing the substrate peptide cleaved from 
      the solid substrate by matrix metalloproteinase-3 (MMP-3). We successfully used 
      this assay to determine the quantitative proteolytic activity of MMP-3 in a 
      dose-dependent manner. In addition, parameters including Michaelis constant 
      (K(m)), maximum rate of enzymatic reaction (V(max)), and half maximal inhibitory 
      concentration (IC(50)) were determined by analyzing the concentrations of 
      substrate peptide cleaved by MMP-3. Therefore, this new assay has potential for 
      the quantitative analysis of enzyme kinetics of protease and informs research 
      developments in drug discovery utilizing kinetic studies.
CI  - Copyright (c) 2012 Elsevier B.V. All rights reserved.
FAU - Kong, Deok-Hoon
AU  - Kong DH
AD  - Department of Molecular and Cellular Biochemistry, Kangwon National University 
      School of Medicine, Chuncheon, Kangwon-Do, Republic of Korea.
FAU - Jung, Se-Hui
AU  - Jung SH
FAU - Lee, Seung-Taek
AU  - Lee ST
FAU - Kim, Young-Myeong
AU  - Kim YM
FAU - Ha, Kwon-Soo
AU  - Ha KS
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120420
PL  - England
TA  - Biosens Bioelectron
JT  - Biosensors & bioelectronics
JID - 9001289
RN  - 0 (5-carboxytetramethylrhodamine succinimidyl ester)
RN  - 0 (Peptides)
RN  - 0 (Rhodamines)
RN  - EC 3.4.24.17 (MMP3 protein, human)
RN  - EC 3.4.24.17 (Matrix Metalloproteinase 3)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Catalysis
MH  - Catalytic Domain
MH  - Cysteine/chemistry
MH  - Drug Discovery
MH  - Fluorescence
MH  - Humans
MH  - Hydrolysis
MH  - Kinetics
MH  - Matrix Metalloproteinase 3/*chemistry
MH  - Peptides/*chemistry
MH  - Phase Transition
MH  - Rhodamines/*chemistry
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
MH  - Substrate Specificity
EDAT- 2012/05/01 06:00
MHDA- 2012/10/26 06:00
CRDT- 2012/05/01 06:00
PHST- 2011/12/17 00:00 [received]
PHST- 2012/03/18 00:00 [revised]
PHST- 2012/04/09 00:00 [accepted]
PHST- 2012/05/01 06:00 [entrez]
PHST- 2012/05/01 06:00 [pubmed]
PHST- 2012/10/26 06:00 [medline]
AID - S0956-5663(12)00220-5 [pii]
AID - 10.1016/j.bios.2012.04.004 [doi]
PST - ppublish
SO  - Biosens Bioelectron. 2012 Jun-Jul;36(1):147-53. doi: 10.1016/j.bios.2012.04.004. 
      Epub 2012 Apr 20.