Real time imaging of calcium-induced localized proteolytic activity after axotomy and its relation to growth cone formation.
AuthorsGitler D, Spira ME
JournalNeuron
PubMed ID9655501
'The emergence of a neuronal growth cone from a transected axon is a necessary step in the sequence of events that leads to successful regeneration. Yet, the molecular mechanisms underlying its formation after axotomy are unknown. In this study, we show by real time imaging of the free intracellular Ca2+ ... More
A homogeneous, nonradioactive high-throughput fluorogenic protein kinase assay.
AuthorsKupcho K, Somberg R, Bulleit B, Goueli SA
JournalAnal Biochem
PubMed ID12758259
Protein kinases play an important role in many cellular processes and mediate cellular responses to a variety of extracellular stimuli. They have been identified by many pharmaceuticals as valid targets for drug discovery. Because of the large number of protein kinases, and the large number of compounds to be screened, ... More
A homogeneous, nonradioactive high-throughput fluorogenic protein phosphatase assay.
AuthorsKupcho K, Hsiao K, Bulleit B, Goueli SA
JournalJ Biomol Screen
PubMed ID15140384
Protein phosphatases are critical components in cellular regulation; they do not only act as antioncogenes by antagonizing protein kinases, but they also play a positive regulatory role in a variety of cellular processes that require dephosphorylation. Thus, assessing the function of these enzymes necessitates the need for a robust, sensitive ... More
Flow cytometric analysis of protease activities in vital cells.
AuthorsRothe G, Klingel S, Assfalg-Machleidt I, Machleidt W, Zirkelbach C, Banati RB, Mangel WF, Valet G
JournalBiol Chem Hoppe Seyler
PubMed ID1381188
The analysis of lysosomal proteases in cell lysates is complicated by pH-dependent and oxidative changes of their activity and complex formation with cytosolic inhibitors. Therefore, new flow cytometric methods were developed for the intracellular measurement of protease activities in viable cells. Intracellular cleavage of substrates such as Z-Arg-Arg-4-trifluoromethylcoumarinyl-7-amide to green ... More