Rabbit Anti-Human PAK1 (Phospho-Thr212) polyclonal antibody

To elucidate the potential function of miRNA-221 in gestational diabetes mellitus (GDM) and the underlying mechanism. MiRNA-221 level was analyzed in the microarray containing placental tissues of GDM rats. After constructing GDM model in rats, miRNA-221 level in placental tissues of GDM rats or controls was determined as well. The relationship between miRNA-221 level and blood glucose in GDM rats was analyzed by Spearman correlation test. Regulatory effects of miRNA-221 on proliferation, apoptosis and insulin secretion in INS-1 cells were assessed. Through dual-luciferase reporter gene assay, the direct target of miRNA-221, PAK1 was identified. At last, potential influences of miRNA-221/PAK1 axis on INS-1 cell phenotypes were determined. MiRNA-221 was downregulated in placental tissues of GDM rats, and its level was negatively correlated to that of blood glucose level in GDM rats. Overexpression of miRNA-221 stimulated insulin secretion, cell proliferation and suppressed apoptosis in INS-1 cells. Knockdown of miRNA-221 achieved the opposite results. PAK1 was proved as the direct target of miRNA-221. Notably, PAK1 was able to reverse regulatory effects of miRNA-221 on INS-1 cell phenotypes. MiRNA-221 regulates proliferation, apoptosis and insulin secretion in islet beta cells through targeting PAK1, thus protecting GDM-induced islet dysfunction. (C) 2019 Elsevier Inc. All rights reserved.

p21-activated kinases (Paks) play an integral component in various cellular diverse processes. The full activation of Pak is dependent upon several serine residues present in the N-terminal region, a threonine present at the activation loop, and finally the phosphorylation of these residues ensure the complete activation of Pak1. The present study deals with the identification of novel potent candidates of Pak1 using computational methods as anti-cancer compounds. A diverse energy based pharmacophore (e-pharmacophore) was developed using four co-crystal inhibitors of Pak1 having pharmacophore features of 5 (DRDRR), 6 (DRHADR), and 7 (RRARDRP and DRRDADH) hypotheses. These models were used for rigorous screening against e-molecule database. The obtained hits were filtered using ADME/T and molecular docking to identify the high affinity binders. These hits were subjected to hierarchical clustering using dendritic fingerprint inorder to identify structurally diverse molecules. The diverse hits were scored against generated water maps to obtain WM/MM Delta G binding energy. Furthermore, molecular dynamics simulation and density functional theory calculations were performed on the final hits to understand the stability of the complexes. Five structurally diverse novel Pak1 inhibitors (4835785, 32198676, 32407813, 76038049, and 32945545) were obtained from virtual screening, water thermodynamics and WM/MM Delta G binding energy. All hits revealed similar mode of binding pattern with the hinge region residues replacing the unstable water molecules in the binding site. The obtained novel hits could be used as a platform to design potent drugs that could be experimentally tested against cancer patients having increased Pak1 expression.