International Journal of Peptides / 2016 / Article / Tab 2

Research Article

Netrin-1 Peptide Is a Chemorepellent in Tetrahymena thermophila

Table 2

Pharmacological studies suggest that a tyrosine kinase is involved in the avoidance of netrin-1 peptide in Tetrahymena. Inhibition of G-proteins, chelation of intracellular and extracellular calcium, and inhibition of a number of other kinases had no effect on avoidance behavior. The tyrosine kinase inhibitor, genistein, reduced avoidance to baseline levels. “Avoidance” represents the mean ± standard deviation of 6 trials. Each trial consisted of 10 cells which were individually observed and scored for avoidance within the first 5 seconds of exposure to netrin-1 peptide.

Pharmacological agentMechanism of actionConcentration testedAvoidance

EGTAChelates extracellular calcium100 μM100 ± 0%
BAPTA-AMChelates intracellular calcium100 μM96.6 ± 5.8%
ThapsigarginDepletes ER calcium stores100 μM100 ± 0%
GDP--SInhibits G-proteins1 mM93.3 ± 5.8%
Pertussis toxinInhibits Gi/o proteins0.2 μg/mL96.6 ± 5.8%
Rp-cAMPsInhibits protein kinase A100 μM93.3 ± 5.8%
U73122Inhibits phospholipase C1 μM95.0 ± 5.0%
GenisteinInhibits tyrosine kinases100 μg/mL16.7 ± 4.5%
DaidzeinNegative control for genistein100 μg/mL98.3 ± 4.1%
Neomycin sulfateCompetitive inhibitor for lysozyme/PACAP receptor100 μM93.3 ± 5.8%
SU6668Protein kinase Inhibitor100 μM100 ± 0%
NS2028Guanylyl cyclase inhibitor100 μM100 ± 0%
GSK429286Rho kinase inhibitor100 μM100 ± 0%
Src inhibitor 1Inhibits Src family kinases100 μM100 ± 0%
FAK inhibitor 14Inhibits focal adhesion kinase100 μM93.3 ± 5.8%
Sodium orthovanadateInhibits tyrosine phosphatases100 μM97.5 ± 5.0%

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