Abstract
The major intracellular protein tyrosine phosphatase (PTP1B) is a 50kDa protein, localized to the endoplasmic reticulum. This PTP is recovered in the particulate fraction of mammalian cells and can be solubilized as a complex of 150 kDa by extraction with non-ionic detergents. Previous work from this laboratory implicated phosphorylation of serine/ threonine residues in the regulation of this PTP. Activity was several-fold higher in cells treated with activators of cAMP-dependent or Ca2+/phospholipid-dependent protein kinases or inhibitors of protein phosphatase 2A. Here we show that these treatments result in more than an 8-fold increase in the phosphorylation of the 50kDa PTP catalytic subunit within the 150 kDa form of the phosphatase in HeLa cells. The phosphorylation occurred exclusively on serine residues, and the same tryptic and cyanogen bromide 32P-phosphopeptides were recovered in the PTP from control and stimulated cells. Either multiple kinases phosphorylate a common site in the PTP1B, or a single kinase is activated ‘downstream’ of cAMP- and Ca2+/phospholipid-dependent kinases. The results indicate that phosphorylation of a serine residue in the segment 283–364, probably serine 352 in the sequence Lys-Gly-Ser-Pro-Leu, occurs in response to cell stimulation. Phosphorylation in this region of PTP1B, between the N-terminal catalytic domain and the C-terminal membrane localization segment, is proposed to regulate phosphatase activity. (Mol Cell Biochem 127/128: 121–129, 1993)
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References
Bishop JM: Molecular themes in oncogenesis. Cell 64: 235–248, 1991
Cantley LC, Auger KR, Carpenter C. Duckworth B, Graziani A, Kapeller R, Soltoff S: Oncogenes and signal transduction. Cell 64: 281–302, 1991
Brautigan DL: Great expectations: protein tyrosine phosphatases in cell regulation. Biochem Biophys Acta 1114: 63–77, 1992
Charbonneau H, Tonks NK: 1002 protein phosphatases? Annu Rev Cell Biol 8: 463–193,1992
Trowbridge IS, Ostergaard HL, Johnson P: CD45: a leukocyte-specific member of the protein tyrosine phosphatase family. Biochim Biophys Acta 1095: 46–56, 1991
Omary MB, Trowbridge IS: Disposition of T200 glycoprotein in the plasma membrane of a murine lymphoma cell line. J Biol Chem 255: 1662–1669, 1980
Valentine MA, Widmer MB, Ledbetter JA, Pinault FM, Voice R, Clark EA, Gallis B, Brautigan DL: Interleukin 2 stimulates serine phosphorylation of CD45 in CTLL-2.4 cells. Eur J Immunol 21: 913–919, 1991
Tonks NK, Diltz CD, Fischer EH: CD45, an integral membrane protein tyrosine phosphatase. J Biol Chem 265: 10674–10680, 1990
Ostergaard HL, Trowbridge IS: Negative regulation of CD45 pro tein tyrosine phosphatase activity by ionomycin in T cells. Science 253: 1423–1425, 1991
Stover DR, Charbonneau H, Tonks NK, Walsh KA: Protein-tyrosine-phosphatase CD45 is phosphorylated transiently on tyrosine upon activation of Jurkat T cells. Proc Natl Acad Sci USA 88: 7704–7707, 1991
Pot DA, Woodford TA, Remboutsika E, Haun RS, Dixon JE: Cloning, bacterial expression, purification, and characterization of the cytoplasmic domain of rat LAR, a receptor-like protein ty rosine phosphatase. J Biol Chem 266: 19688–19696, 1991
Feng GS, Hui CC, Pawson T: SH2-containing phosphotyrosine phosphatase as a target of protein-tyrosine kinases. Science 259: 1607–1611, 1993
Vogel W, Lammers R, Huang J, Ullrich A: Activation of a phosphotyrosine phosphatase by tyrosine phosphorylation. Science 259: 1611–1614, 1993
Rotenberg SA, Brautigan DL: Membrane protein phosphotyrosine phosphatase in rabbit kidney. Biochem J 243: 747–754, 1987
Charbonneau H, Tonks NK, Kumar S, Diltz CD, Harrylock M, Cool DE, Krebs EG. Fischer EH, Walsh KA: Human placenta protein-tyrosine-phosphatase: amino acid sequence and relationship to a family of receptor-like proteins. Proc Natl Acad Sci USA 86: 5252–5256, 1989
Woodford-Thomas TA, Rhodes JD, Dixon JE: Expression of a protein tyrosine phosphatase in normal and v-src-transformed mouse 3T3 fibroblasts. J Cell Biol 117: 401–414, 1992
Frangioni JV, Beahm PH, Shifrin V, Jost CA, Neel BG: The nontransmembrane tyrosine phosphatase PTP-1B localizes to the endoplasmic reticulum via its 35 amino acid c-terminal sequence. Cell 68: 545–560, 1992
Brautigan DL, Pinault FM: Activation of membrane protein-tyrosine phosphatase involving cAMP and Ca2+/phospholipid-dependent protein kinases. Proc Natl Acad Sci USA 88 (15): 6696–6700, 1991
Shriner CL, Brautigan DL: Cytosolic protein phosphotyrosine phosphatase from rabbit kidney. J Biol Chem 259 (18): 11383–11390, 1984
Cool DE, Tonks NK, Charbonneau H. Fischer EH, Krebs EG: Expression of a human T-cell protein-tyrosine-phosphatase in baby hamster kidney cells. Proc Natl Acad Sci USA 87: 7280–7284, 1990
Pallen CJ, Tong PH: Elevation of membrane tyrosine phosphatase activity in density-dependent growth-arrested fibroblasts. Proc Natl Acad Sci USA 88: 6996–7000. 1991
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© 1993 Kluwer Academic Publishers
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Brautigan, D.L., Pinault, F.M. (1993). Serine phosphorylation of protein tyrosine phosphatase (PTP1B) in HeLa cells in response to analogues of cAMP or diacylglycerol plus okadaic acid. In: Khandelwal, R.L., Wang, J.H. (eds) Reversible Protein Phosphorylation in Cell Regulation. Developments in Molecular and Cellular Biochemistry, vol 11. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2600-1_11
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DOI: https://doi.org/10.1007/978-1-4615-2600-1_11
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