A cation counterflux supports lysosomal acidification
Open Access
- 21 June 2010
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 189 (7), 1171-1186
- https://doi.org/10.1083/jcb.200911083
Abstract
Protein interactions are involved in all cellular processes. Their efficient and reliable characterization is therefore essential for understanding biological mechanisms. In this study, we show that combining bacterial artificial chromosome (BAC) TransgeneOmics with quantitative interaction proteomics, which we call quantitative BAC–green fluorescent protein interactomics (QUBIC), allows specific and highly sensitive detection of interactions using rapid, generic, and quantitative procedures with minimal material. We applied this approach to identify known and novel components of well-studied complexes such as the anaphase-promoting complex. Furthermore, we demonstrate second generation interaction proteomics by incorporating directed mutational transgene modification and drug perturbation into QUBIC. These methods identified domain/isoform-specific interactors of pericentrin- and phosphorylation-specific interactors of TACC3, which are necessary for its recruitment to mitotic spindles. The scalability, simplicity, cost effectiveness, and sensitivity of this method provide a basis for its general use in small-scale experiments and in mapping the human protein interactome.Keywords
This publication has 43 references indexed in Scilit:
- Revisiting the Role of Cystic Fibrosis Transmembrane Conductance Regulator and Counterion Permeability in the pH Regulation of Endocytic OrganellesMolecular Biology of the Cell, 2009
- Burkholderia cenocepacia-induced delay of acidification and phagolysosomal fusion in cystic fibrosis transmembrane conductance regulator (CFTR)-defective macrophagesMicrobiology, 2008
- The Cl-/H+ antiporter ClC-7 is the primary chloride permeation pathway in lysosomesNature, 2008
- Cystic Fibrosis Transmembrane Conductance Regulator-independent Phagosomal Acidification in MacrophagesOnline Journal of Public Health Informatics, 2007
- Pannexin-1 mediates large pore formation and interleukin-1β release by the ATP-gated P2X7 receptorThe EMBO Journal, 2006
- ClC-7 requires Ostm1 as a β-subunit to support bone resorption and lysosomal functionNature, 2006
- A small molecule CFTR inhibitor produces cystic fibrosis‐like submucosal gland fluid secretions in normal airwaysThe FASEB Journal, 2004
- Thiazolidinone CFTR inhibitor identified by high-throughput screening blocks cholera toxin–induced intestinal fluid secretionJCI Insight, 2002
- Ionophore Properties of Monensin Derivatives Studied on Human Erythrocytes by 23Na NMR and K+ and H+ Potentiometry: Relationship with Antimicrobial and Antimalarial ActivitiesJournal of Medicinal Chemistry, 1996
- Potassium ion dependent proton efflux and depolarization from spleen lysosomesBiochemical and Biophysical Research Communications, 1988