title: Ubiquitin receptor binding and signaling in primary human leukocytes.
abstract: Utilizing the human monocyte/macrophage cell line THP1, we recently identified extracellular ubiquitin as an endogenous agonist of the G protein-coupled receptor CXC chemokine receptor (CXCR) 4. Because receptor binding and signaling properties of extracellular ubiquitin have not been evaluated in primary human leukocytes, we analyzed its binding characteristics and subsequent Ca(2+) signaling in freshly isolated human B cells, T cells and monocytes. Ubiquitin binding shows typical receptor binding characteristics and promotes intracellular Ca(2+) flux within seconds in all three cell populations. The K(d) for the ubiquitin receptor interaction in freshly isolated human monocytes is consistent with the affinity of the ubiquitin CXCR4 interaction that we reported for THP1 cells. As detected in THP1 cells previously, the ubiquitin induced Ca(2+) flux can be attenuated with a phospholipase C inhibitor in all primary leukocyte cultures. Our observations further support the finding that ubiquitin is a CXCR4 agonist and demonstrate that extracellular ubiquitin induces physiological relevant signaling events in primary human leukocytes. Although the exact mechanism of the ubiquitin CXCR4 interaction, its receptor selectivity and subsequent signaling events remain to be determined, our findings identify a novel and unexpected biological role of extracellular ubiquitin as an endogenous immune modulator.
authors.affiliation: Burn and Shock Trauma Institute; Department of Surgery; Loyola University; Chicago Stritch School of Medicine; Maywood, IL USA.
authors.forename: Vikas
authors.initials: V
authors.lastname: Saini
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Romero
authors.forename: Adriano
authors.initials: A
authors.lastname: Marchese
authors.forename: Matthias
authors.initials: M
authors.lastname: Majetschak