Two Novel Peripheral Membrane Proteins, Pasin 1 and Pasin 2, Associated with Na+,K+-ATPase in Various Cells and Tissues

• Published in: The Journal of cell biology 1990-12, Vol.111 (6), p.2375-2383
• Main Authors: Kraemer, Doris, Koob, Rainer, Friedrichs, Bärbel, Drenckhahn, Detlev
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Antibodies; Biological and medical sciences; Brain - enzymology; Cell Membrane - enzymology; Cell membranes; Electrophoresis, Polyacrylamide Gel; Epithelial cells; Erythrocyte membrane; Erythrocytes; Fundamental and applied biological sciences. Psychology; Gels; Kidney - enzymology; Kidney Medulla - enzymology; Kidneys; Membrane proteins; Membrane Proteins - isolation & purification; Microsomes; Miscellaneous; Molecular Sequence Data; Molecular Weight; Organ Specificity; Ouabain - pharmacology; P branes; Parotid Gland - enzymology; Peptide Fragments - isolation & purification; Peptide Mapping; Proteins; Sodium-Potassium-Exchanging ATPase - isolation & purification; Sodium-Potassium-Exchanging ATPase - metabolism; Swine
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.111.6.2375

Purification of pig kidney Na+,K+-ATPase at low concentrations of SDS (0.5%) allowed copurification of several peripheral membrane proteins. Some of these associated proteins were identified as components of the membrane cytoskeleton. Here we describe two novel globular proteins of of Mr 77,000 (pasin 1) and Mr 73,000 (pasin 2) which copurify and coimmunoprecipitate with Na+,K+-ATPase and can be stripped off Na+,K+-ATPase microsomes by 1 M KCl. Pasin 1 and pasin 2 were detected by immunoblot analysis in various cells and tissues including erythrocytes and platelets. Immunostaining revealed colocalization of pasin 1 and Na+,K+-ATPase along the basolateral cell surface of epithelial cells of kidney tubules and parotid striated ducts (titers of pasin 2 antibodies were too weak for immunocytochemistry). In erythrocytes, pasin 1 and pasin 2 are minor components bound to the cytoplasmic surface of the plasma membrane. Pasin 1 showed the same electrophoretic mobility as protein 4.1b. However, both proteins have different isoelectric points (pasin 1, pI 6; protein 4.1, pI 7), different chymotryptic fragments, and are immunologically unrelated. Short pieces of sequence obtained from pasin 1 and pasin 2 were not found in any other known protein sequence. The occurrence of pasin 1 and pasin 2 in diverse cells and tissues and their association with Na+,K+-ATPase suggests a general role of these proteins in Na+,K+-ATPase function.