+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Application of a novel highly sensitive activity-based probe for detection of cathepsin G

Application of a novel highly sensitive activity-based probe for detection of cathepsin G

Analytical Biochemistry 421(2): 667-672

Cathepsins are crucial in antigen processing in the major histocompatibility complex class II (MHC II) pathway. Within the proteolytic machinery, three classes of proteases (i.e., cysteine, aspartic, and serine proteases) are present in the endocytic compartments. The combined action of these proteases generates antigenic peptides from antigens, which are loaded to MHC II molecules for CD4+ T cell presentation. Detection of active serine proteases in primary human antigen-presenting cells (APCs) is restricted because of the small numbers of cells isolated from the peripheral blood. For this purpose, we developed a novel highly sensitive α-aminoalkylphosphonate diphenyl ester (DAP) activity-based probe to detect the serine protease cathepsin G (CatG) in primary APCs and after Epstein-Barr virus (EBV) exposure. Although CatG activity was not altered after short-term exposure of EBV in primary myeloid dendritic cells 1 (mDC1s), the aspartic protease cathepsin D (CatD) was reduced, suggesting that EBV is responsible for mitigating the presentation of a model antigen tetanus toxoid C-fragment (TTCF) by reduction of CatD. In addition, CatG activity was reduced to background levels in B cells during cell culture; however, these findings were independent of EBV transformation. In conclusion, our activity-based probe can be used for both Western blot and 96-well-based high-throughput CatG detection when cell numbers are limited.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 051607527

Download citation: RISBibTeXText

PMID: 22178917

DOI: 10.1016/j.ab.2011.11.016

Related references

Application of a novel high sensitive activity-based probe for detection of cathepsin G. 2011

Design of a highly selective quenched activity-based probe and its application in dual color imaging studies of cathepsin S activity localization. Journal of the American Chemical Society 137(14): 4771-4777, 2015

A highly sensitive G-quadruplex-based luminescent switch-on probe for the detection of polymerase 3? to 5? proofreading activity. 2013

A highly sensitive G-quadruplex-based luminescent switch-on probe for the detection of polymerase 3'-5' proofreading activity. Methods 64(3): 224-228, 2013

Highly sensitive protein detection based on a novel probe with catalytic activity combined with a signal amplification strategy: assay of MDM2 for cancer staging. Chemical Communications 49(84): 9848-9850, 2013

A highly sensitive and facile graphene oxide-based nucleic acid probe: Label-free detection of telomerase activity in cancer patient's urine using AIEgens. Biosensors and Bioelectronics 89(Pt 1): 417-421, 2017

Naked-eye Detection of Hg 2+ in Practical Applications Using a Highly Selective and Sensitive Fluorescent Probe. Analytical Sciences 34(12): 1411-1417, 2018

Highly Sensitive Fluorescence Probe Based on Functional SBA-15 for Selective Detection of Hg. Nanoscale Research Letters 5(9): 1468-1473, 2010

A highly selective and sensitive probe for Cu2+ based on rhodamine-pyridazine conjugate and its application. Journal of Fluorescence 25(1): 15-24, 2015

A highly sensitive upconverting phosphors-based offon probe for the detection of glutathione. Sensors and Actuators B: Chemical 185: 363-369, 2013

Highly sensitive iridium(iii) complex-based phosphorescent probe for thiophenol detection. Dalton Transactions 46(39): 13456-13462, 2017

Highly Sensitive and Selective Fluorescent Probe for Detection of Fe 3+ Based on Rhodamine Fluorophore. Journal of Fluorescence 29(3): 645-652, 2019

Branch-Migration Based Fluorescent Probe for Highly Sensitive Detection of Mercury. Analytical Chemistry 90(20): 11764-11769, 2018

A gold nanoparticle-based fluorescence turn-on probe for highly sensitive detection of polyamines. Chemistry 17(43): 11978-11982, 2011

Highly sensitive rare cell detection based on quantum dot probe fluorescence analysis. Analytical and Bioanalytical Chemistry 396(3): 1135-1141, 2010