title: Kinase activity profiling of gram-negative pneumonia.
abstract: Pneumonia is a severe disease with high morbidity and mortality. A major causative pathogen is the Gram-negative bacterium Klebsiella (K.) pneumoniae. Kinases play an integral role in the transduction of intracellular signaling cascades and regulate a diverse array of biological processes essential to immune cells. The current study explored signal transduction events during murine Gram-negative pneumonia using a systems biology approach. Kinase activity arrays enable the analysis of 1,024 consensus sequences of protein kinase substrates. Using a kinase activity array on whole lung lysates, cellular kinase activities were determined in a mouse model of K. pneumoniae pneumonia. Notable kinase activities also were validated with phospho-specific Western blots. On the basis of the profiling data, mitogen-activated protein kinase (MAPK) signaling via p42 mitogen-activated protein kinase (p42) and p38 mitogen-activated protein kinase (p38) and transforming growth factor β (TGFβ) activity were reduced during infection, whereas v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian) (SRC) activity generally was enhanced. AKT signaling was represented in both metabolic and inflammatory (mitogen-activated protein kinase kinase 2 [MKK], apoptosis signal-regulating kinase/mitogen-activated protein kinase kinase kinase 5 [ASK] and v-raf murine sarcoma viral oncogene homolog B1 [b-RAF]) context. This study reaffirms the importance of classic inflammation pathways, such as MAPK and TGFβ signaling and reveals less known involvement of glycogen synthase kinase 3β (GSK-3β), AKT and SRC signaling cassettes in pneumonia.
authors.affiliation: Center for Infection and Immunity Amsterdam, The Netherlands. a.j.hoogendijk@amc.uva.nl
authors.forename: Arie J
authors.initials: AJ
authors.lastname: Hoogendijk
authors.forename: Sander H
authors.initials: SH
authors.lastname: Diks
authors.forename: Maikel P
authors.initials: MP
authors.lastname: Peppelenbosch
authors.forename: Tom
authors.initials: T
authors.lastname: Van Der Poll
authors.forename: Catharina W
authors.initials: CW
authors.lastname: Wieland
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018925:Chemokines / Q000378:metabolism; D016000:Cluster Analysis; D016207:Cytokines / Q000378:metabolism; D005260:Female; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D054884:Host-Pathogen Interactions; D007710:Klebsiella Infections / Q000201:enzymology* / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000502:physiology; D008168:Lung / Q000201:enzymology / Q000378:metabolism / Q000382:microbiology; D020935:MAP Kinase Signaling System; D051379:Mice; D008810:Mice, Inbred C57BL; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D010766:Phosphorylation; D010770:Phosphotransferases / Q000145:classification / Q000378:metabolism*; D018410:Pneumonia, Bacterial / Q000201:enzymology* / Q000382:microbiology; D040901:Proteomics / Q000379:methods*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D018925:Chemokines; D016207:Cytokines; D016212:Transforming Growth Factor beta; D010770:Phosphotransferases; D019061:src-Family Kinases; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D051057:Proto-Oncogene Proteins c-akt; D019950:Mitogen-Activated Protein Kinase 1; D048051:p38 Mitogen-Activated Protein Kinases; D038362:Glycogen Synthase Kinase 3