title: Phage display based cloning of proteins interacting with the cytoplasmic tail of membrane immunoglobulins.
abstract: The reduced quantity and quality of serum immunoglobulins (sIgs) in mutant mice expressing truncated cytoplasmic tails of IgE and IgG1 indicate an active role for the cytoplasmic domains of mIgG1 and mIgE. We used phage display technology to identify candidate proteins able to interact with the cytoplasmic tail of mIgE. Using a murine cDNA B cell library displayed on the surface of phage as prey and the 28 amino acid long cytoplasmic tail of IgE as bait, we isolated phage encoding the murine hematopoietic progenitor kinase 1 (HPK1). Surface plasmon resonance analysis measurements confirmed affinity of HPK1 to the mIgE cytoplasmic tail and revealed association to other immunoglobulin isotypes as well. Immunoprecipitation experiments, using lysates from two B cell lines expressing nitrophenyl (NP) specific mIgE molecules showed co-precipitation of IgE and HPK1. The interaction of HPK1 with the cytoplasmic domains of membrane immunoglobulins indicate an active role of the tails as part of an isotype specific signal transduction, independent from the Igalpha/Igbeta heterodimers, and may represent a missing link to upstream regulatory elements of HPK1 activation.
authors.affiliation: Department of Genetics and General Biology, Hellbrunnerstrasse 34, A-5020 Salzburg, Austria.
authors.forename: Roland
authors.initials: R
authors.lastname: Geisberger
authors.forename: Martin
authors.initials: M
authors.lastname: Prlic
authors.forename: Gertrude
authors.initials: G
authors.lastname: Achatz-Straussberger
authors.forename: Iris
authors.initials: I
authors.lastname: Oberndorfer
authors.forename: Elke
authors.initials: E
authors.lastname: Luger
authors.forename: Marinus
authors.initials: M
authors.lastname: Lamers
authors.forename: Reto
authors.initials: R
authors.lastname: Crameri
authors.forename: Ulrich
authors.initials: U
authors.lastname: Appenzeller
authors.forename: Jürgen
authors.initials: J
authors.lastname: Wienands
authors.forename: Michael
authors.initials: M
authors.lastname: Breitenbach
authors.forename: Fatima
authors.initials: F
authors.lastname: Ferreira
authors.forename: Gernot
authors.initials: G
authors.lastname: Achatz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D003001:Cloning, Molecular; D018076:DNA, Complementary; D007073:Immunoglobulin E / Q000737:chemistry / Q000378:metabolism; D007132:Immunoglobulin Isotypes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D010954:Plasmacytoma; D011233:Precipitin Tests; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011947:Receptors, Antigen, B-Cell / Q000737:chemistry / Q000378:metabolism*; D017455:Receptors, IgE / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020349:Surface Plasmon Resonance; D014407:Tumor Cells, Cultured
chemical_list: D018076:DNA, Complementary; D007132:Immunoglobulin Isotypes; D019151:Peptide Library; D010455:Peptides; D011506:Proteins; D011947:Receptors, Antigen, B-Cell; D017455:Receptors, IgE; D007073:Immunoglobulin E; C102102:hematopoietic progenitor kinase 1; D017346:Protein Serine-Threonine Kinases