title: Isolation of serpin-interacting proteins in C. elegans using protein affinity purification.
abstract: Caenorhabditis elegans is a useful model organism for combining multiple imaging, genetic, and biochemical methodologies to gain more insight into the biological function of specific proteins. Combining both biochemical and genetic analyses can lead to a better understanding of how a given protein may function within the context of a network of other proteins or specific pathway. Here, we describe a protocol for the biochemical isolation of serpin-interacting proteins using affinity purification and proteomic analysis. As the knowledge of in vivo serpin interacting partners in C. elegans has largely been obtained using genetic and in vitro recombinant protein studies, this protocol serves as a complementary approach to provide insight into the biological function and regulation of serpins.
authors.affiliation: Department of Pediatrics, Cell Biology and Physiology, University of Pittsburgh School of Medicine, Children's Hospital of Pittsburgh of UPMC, and Magee-Womens Hospital of UPMC, Pittsburgh, PA 15224, USA.
authors.forename: Mark T
authors.initials: MT
authors.lastname: Miedel
authors.affiliation: Biomedical Mass Spectrometry Center, University of Pittsburgh Schools of the Health Sciences, Pittsburgh, PA 15213, USA.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Zeng
authors.affiliation: Biomedical Mass Spectrometry Center, University of Pittsburgh Schools of the Health Sciences, Pittsburgh, PA 15213, USA; Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Yates
authors.affiliation: Department of Pediatrics, Cell Biology and Physiology, University of Pittsburgh School of Medicine, Children's Hospital of Pittsburgh of UPMC, and Magee-Womens Hospital of UPMC, Pittsburgh, PA 15224, USA.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Silverman
authors.affiliation: Department of Pediatrics, Cell Biology and Physiology, University of Pittsburgh School of Medicine, Children's Hospital of Pittsburgh of UPMC, and Magee-Womens Hospital of UPMC, Pittsburgh, PA 15224, USA. Electronic address: cliff.luke@chp.edu.
authors.forename: Cliff J
authors.initials: CJ
authors.lastname: Luke
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D011485:Protein Binding; D060066:Protein Interaction Maps / Q000235:genetics; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D015843:Serpins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D029742:Caenorhabditis elegans Proteins; D011994:Recombinant Proteins; D015843:Serpins
keywords: Affinity purification; Caenorhabditis elegans; Proteomics; Serpin