title: The gap-junction protein connexin 56 is phosphorylated in the intracellular loop and the carboxy-terminal region.
abstract: The lens gap-junction protein, connexin 56, is modified by phosphorylation. Two-dimensional mapping of tryptic phosphopeptides of 32P-labeled connexin 56 from primary chicken-lens cultures showed that treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA) induced an increase in phosphorylation of connexin 56 at specific constitutively phosphorylated sites. Treatment with 8-Br-cAMP or forskolin did not induce substantial changes in connexin 56 phosphorylation. Two phosphorylation sites within connexin 56, S493 and S118, were identified after HPLC purification and peptide sequencing of tryptic phosphopeptides from bacterially expressed connexin 56 fusion proteins phosphorylated by protein kinase C or protein kinase A in vitro. Comparisons of the two-dimensional maps of tryptic phosphopeptides from in vitro phosphorylated connexin 56 fusion proteins and in vivo phosphorylated connexin 56 showed that S493 and S118 were constitutively phosphorylated in lentoid-containing cultures, and that treatment with TPA induced an increase in phosphorylation of the peptides containing S118. It is suggested that phosphorylation of connexin 56 at S118 is involved in the TPA-induced decrease in intercellular communication and acceleration of connexin 56 degradation.
authors.affiliation: Department of Pediatrics, Washington University School of Medicine, St Louis, MO 63110, USA.
authors.forename: V M
authors.initials: VM
authors.lastname: Berthoud
authors.forename: E C
authors.initials: EC
authors.lastname: Beyer
authors.forename: W E
authors.initials: WE
authors.lastname: Kurata
authors.forename: A F
authors.initials: AF
authors.lastname: Lau
authors.forename: P D
authors.initials: PD
authors.lastname: Lampe
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002642:Chick Embryo; D017630:Connexins / Q000302:isolation & purification / Q000378:metabolism*; D005136:Eye Proteins / Q000302:isolation & purification / Q000378:metabolism*; D017629:Gap Junctions / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D007908:Lens, Crystalline / Q000737:chemistry; D009924:Organ Culture Techniques; D010446:Peptide Fragments / Q000032:analysis; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014357:Trypsin
chemical_list: D017630:Connexins; D005136:Eye Proteins; D010446:Peptide Fragments; D010761:Phosphorus Radioisotopes; D011993:Recombinant Fusion Proteins; C078866:connexin 56; D005982:Glutathione Transferase; D014357:Trypsin