English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Probing the kinetics of quantum dot-based proteolytic sensors.

Diaz, S. A., Malonoski, A. P., Susumu, K., Hofele, R. V., Oh, E., & Medintz, I. L. (2015). Probing the kinetics of quantum dot-based proteolytic sensors. Analytical and Bioanalytical Chemistry, 407(24), 7307-7318. doi:10.1007/s00216-015-8892-y.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-8982-D Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-6070-8
Genre: Journal Article

Files

show Files
hide Files
:
2214162.pdf (Publisher version), 4MB
 
File Permalink:
-
Name:
2214162.pdf
Description:
-
Visibility:
Restricted (Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute), Göttingen; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
2214162_Suppl.pdf (Supplementary material), 2MB
Name:
2214162_Suppl.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Diaz, S. A., Author
Malonoski, A. P., Author
Susumu, K., Author
Hofele, R. V.1, Author              
Oh, E., Author
Medintz, I. L., Author
Affiliations:
1Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

Content

show
hide
Free keywords: Proteases; Quantumdot; FRET; Enzyme; Sensor; Michaelis-Menten
 Abstract: As an enzyme superfamily, proteases are rivaled only by kinases in terms of their abundance within the human genome. Two ratiometric quantum dot (QD) Forster resonance energy transfer-based sensors designed to monitor the activity of the proteolytic enzymes collagenase and elastase are investigated here. Given the unique material constraints of these sensing constructs, assays are realized utilizing excess enzyme and fixed substrate in progress curve format to yield enzyme specificity or k (cat)/K (m) ratios. The range of k (cat)/K-m values derived is 0.5-1.1 mM(-1) s(-1) for the collagenase sensor and 3.7-4.2 mM(-1) s(-1) for the elastase sensor. Of greater interest is the observation that the elastase sensor can be well represented by the Michaelis-Menten model while the collagenase sensor cannot. The latter demonstrates increased specificity at higher peptide substrate/QD loading values and an apparent QD-caused reversible inhibition as the reaction progresses. Understanding the detailed kinetic mechanisms that underpin these types of sensors will be important especially for their further quantitative utilization.

Details

show
hide
Language(s): eng - English
 Dates: 2015-07-282015-09
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1007/s00216-015-8892-y
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Analytical and Bioanalytical Chemistry
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 407 (24) Sequence Number: - Start / End Page: 7307 - 7318 Identifier: -