Protein Profiling of Active Cysteine Cathepsins in Living Cells Using an Activity-Based Probe Containing a Cell-Penetrating Peptide

2012 ◽  
Vol 11 (12) ◽  
pp. 5763-5772 ◽  
Author(s):  
Fengkai Fan ◽  
Si Nie ◽  
Eric B. Dammer ◽  
Duc M. Duong ◽  
Deng Pan ◽  
...  
Author(s):  
Florina Zakany ◽  
Mate Szabo ◽  
Gyula Batta ◽  
Levente Kárpáti ◽  
István M. Mándity ◽  
...  

Although the largely positive intramembrane dipole potential (DP) may substantially influence the function of transmembrane proteins, its investigation is deeply hampered by the lack of measurement techniques suitable for high-throughput examination of living cells. Here, we describe a novel emission ratiometric flow cytometry method based on F66, a 3-hydroxiflavon derivative, and demonstrate that 6-ketocholestanol, cholesterol and 7-dehydrocholesterol, saturated stearic acid (SA) and ω-6 γ-linolenic acid (GLA) increase, while ω-3 α-linolenic acid (ALA) decreases the DP. These changes do not correlate with alterations in cell viability or membrane fluidity. Pretreatment with ALA counteracts, while SA or GLA enhances cholesterol-induced DP elevations. Furthermore, ALA (but not SA or GLA) increases endo-lysosomal escape of penetratin, a cell-penetrating peptide. In summary, we have developed a novel method to measure DP in large quantities of individual living cells and propose ALA as a physiological DP lowering agent facilitating cytoplasmic entry of penetratin.


2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Makoto Oba ◽  
Takuma Kato ◽  
Kaori Furukawa ◽  
Masakazu Tanaka

2007 ◽  
Vol 18 (4) ◽  
pp. 1325-1331 ◽  
Author(s):  
Adams Amantana ◽  
Hong M. Moulton ◽  
Melissa L. Cate ◽  
Muralimohan T. Reddy ◽  
Tom Whitehead ◽  
...  

2011 ◽  
Vol 112 (9) ◽  
pp. 2454-2462
Author(s):  
Seung Won Yang ◽  
Seung-Min Lee ◽  
Eun Young Choi ◽  
Kyung Hye Lee ◽  
Soo Hyuk Kim ◽  
...  

Biomaterials ◽  
2014 ◽  
Vol 35 (13) ◽  
pp. 4082-4087 ◽  
Author(s):  
Likun Fei ◽  
Li-Peng Yap ◽  
Peter S. Conti ◽  
Wei-Chiang Shen ◽  
Jennica L. Zaro

2020 ◽  
Vol 17 ◽  
pp. 250-256
Author(s):  
Shanna J. Smith ◽  
Caroline M. Li ◽  
Robert G. Lingeman ◽  
Robert J. Hickey ◽  
Yilun Liu ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document