Metabolic 32P Labeling and Purification of IGFBP-5 from T47D Human Breast Carcinoma Cells—
Tryptic Digestion and HPLC—
Alkaline Phosphatase Treatment and Mass Spectrometry—
Phosphorylation in Vitro with CK2—
IGFBP-5 Is Secreted as a Phosphoprotein—
Phosphorylation Sites of IGFBP-5 from T47D Cells—
|Sequence||Fractions||Experimental m/z||Theoretical m/z|
|Monoisotopic, +ve||Average, +ve||Average, −ve|
|30 and 31||2,643.0||2,642.8|
|p100–142||29 and 30||4,384.3||4,383.4|
Glycosylation Sites of IGFBP-5—
Post-translational Modification of Intact IGFBP-5 in Vivo—
|Percentage of total protein|
|28,556, 28,571, and 28,586||0, 15, and 30||None||31.3|
|28,636, 28,651, and 28,666||80, 95, and 110||HPO3||37.2|
|28,716 and 28,734||160 and 178||(HPO3)2||4.1|
|28,839||283||HPO3 + HexNAc||0.8|
|28,922||366||HexNAc and Hex||1.0|
|29,001||444||HPO3 + HexNAc and Hex||1.5|
|29,047||491||HexNAc and NeuAc or HPO3 + (HexNAc)2||0.9|
|29,212 and 29,228||656 and 672||HexNAc, Hex, and NeuAc||2.8|
|29,294 and 29,309||738, 753, and 771||HPO3 + HexNAc, Hex, and NeuAc||3.7|
|29,505, 29,520, and 29,535||949, 964, and 979||HexNAc, Hex, and (NeuAc)2||6.3|
|29,583, 29,602, and 29,617||1,027, 1,046, and 1,061||HPO3 + HexNAc, Hex, and (NeuAc)2||9.7|
Phosphorylation of IGFBP-5 in Vitro with Casein Kinase 2—
Functional Evaluation of IGFBP-5 Phosphorylation and Glycosylation—
- Craig A.G.
- Hoeger C.A.
- Miller C.L.
- Goedken T.
- Rivier J.E.
- Fischer W.H.
- Sala A.
- Capaldi S.
- Campagnoli M.
- Faggion B.
- Labò S.
- Perduca M.
- Romano A.
- Carrizo M.E.
- Valli M.
- Visai L.
- Minchiotti L.
- Galliano M.
- Monaco H.L.
- Imai Y.
- Busby Jr., W.H.
- Smith C.E.
- Clarke J.B.
- Garmong A.J.
- Horwitz G.D.
- Rees C.
- Clemmons D.R.
- Insulin-like growth factor (IGF) binding proteins: Interactions with IGFs and intrinsic bioactivities.Am. J. Physiol. 2000; 278: E967-E976
- Cellular actions of the insulin-like growth factor binding proteins.Endocr. Rev. 2002; 23: 824-854
- Insulin-like growth factor binding protein 5 and apoptosis in mammary epithelial cells.J. Cell Sci. 2003; 116: 675-682
- Human osteoblast-derived insulin-like growth factor (IGF) binding protein-5 stimulates osteoblast mitogenesis and potentiates IGF action.J. Biol. Chem. 1992; 267: 22467-22472
- Evidence that IGF-binding protein-5 functions as a growth factor.J. Clin. Investig. 2001; 107: 73-81
- The effect of phosphorylation by casein kinase 2 on the activity of insulin-like growth factor binding protein.Endocrinology. 2000; 141: 564-570
- Phosphorylation of insulin-like growth factor binding protein-3 by deoxyribonucleic acid-dependent protein kinase reduces ligand binding and enhances nuclear accumulation.Endocrinology. 2003; 144: 1984-1993
- Phosphorylation by DNA-dependent protein kinase is critical for apoptosis induction by insulin-like growth factor binding protein-3.Cancer Res. 2006; 66: 10878-10884
- Insulin-like growth factors and their binding proteins: biological actions.Endocr. Rev. 1995; 16: 3-34
- The role of glycosylation in the action of IGFBP-3.Prog. Growth Factor Res. 1995; 6: 223-229
- Identification of N-linked oligosaccharides of rat insulin-like growth factor binding protein-4.Growth Horm. IGF Res. 2002; 12: 169-177
- Isolation and characterization of circulating 13-kDa C-terminal fragments of human insulin-like growth factor binding protein-5.FEBS Lett. 1998; 441: 281-286
- O-Glycosylation of insulin-like growth factor (IGF) binding protein-6 maintains high IGF-II binding affinity by decreasing binding to glycosaminoglycans and susceptibility to proteolysis.Eur. J. Biochem. 2000; 267: 5378-5386
- Isolation and characterization of a new placenta specific protein (PP12).Arch. Gynecol. 1980; 229: 279-291
- Characterisation of recombinant glycosylation variants of insulin-like growth factor binding protein-3.J. Endocrinol. 1999; 160: 379-387
- Structure of the IGF-binding domain of the insulin-like growth factor-binding protein-5 (IGFBP-5): implications for IGF and IGF-I receptor interactions.EMBO J. 1998; 17: 6558-6572
- Insulin-like growth factor binding protein-3 is secreted as a phosphoprotein by human breast cancer cells.Mol. Cell. Endocrinol. 1999; 156: 131-139
- Mutagenesis of basic amino acids in the carboxyl-terminal region of insulin-like growth factor binding protein-5 affects acid-labile subunit binding.Endocrinology. 2001; 142: 2147-2150
- Multisite phosphorylation of doublecortin by cyclin-dependent kinase 5.Biochem. J. 2004; 381: 471-481
- Reverse staining of sodium dodecyl sulfate polyacrylamide gels by imidazole-zinc salts: sensitive detection of unmodified proteins.BioTechniques. 1992; 12: 564-573
- Improved detection of hydrophilic phosphopeptides using graphite powder microcolumns and mass spectrometry: evidence for in vivo doubly phosphorylated dynamin I and dynamin III.Mol. Cell. Proteomics. 2004; 3: 456-465
- Monitoring protein kinase and phosphatase reactions with matrix-assisted laser desorption/ionization mass spectrometry and capillary zone electrophoresis: comparison of the detection efficiency of peptide-phosphopeptide mixtures.Biol. Mass Spectrom. 1994; 23: 519-528
- Determination of phosphorylation levels of tyrosine hydroxylase by electrospray mass spectrometry.Anal. Biochem. 2000; 281: 98-104
- Protein pI shifts due to posttranslational modifications in the separation and characterization of proteins.Anal. Chem. 2005; 77: 2745-2755
- Identification of the sites of phosphorylation in insulin-like growth factor binding protein-1. Regulation of its affinity by phosphorylation of serine 101.J. Biol. Chem. 1993; 268: 1125-1131
- Identification of the major sites of phosphorylation in IGF binding protein-3.J. Cell. Biochem. 1994; 56: 262-273
- Fibronectin binds insulin-like growth factor-binding protein 5 and abolishes its ligand-dependent action on cell migration.J. Biol. Chem. 2004; 279: 4269-4277
- Insulin-like growth factor binding protein-3 prevents retinoid receptor heterodimerization: implications for retinoic acid-sensitivity in human breast cancer cells.Biochem. Biophys. Res. Commun. 2004; 314: 83-88
- Ras-association domain family 1 protein, RASSF1C, is an IGFBP-5 binding partner and a potential regulator of osteoblast cell proliferation.J. Bone Miner. Res. 2005; 20: 1430-1439
- Insulin-like growth factor-binding protein 5 (IGFBP-5) interacts with a four and a half LIM protein 2 (FHL2).J. Biol. Chem. 2002; 277: 12053-12060
- Characterization of insulin-like growth factor binding protein-1 kinases from human hepatoma cells.J. Cell. Biochem. 1996; 60: 387-399
- Protease-resistant form of insulin-like growth factor-binding protein 5 is an inhibitor of insulin-like growth factor-I actions on porcine smooth muscle cells in culture.J. Clin. Investig. 1997; 100: 2596-2605
- Pregnancy-associated plasma protein-A2 (PAPP-A2), a novel insulin-like growth factor-binding protein-5 proteinase.J. Biol. Chem. 2001; 276: 21849-21853
- Carboxy-truncated insulin-like growth factor binding protein-5 stimulates mitogenesis in osteoblast-like cells.Biochem. Biophys. Res. Commun. 1993; 195: 25-30
- Plasmin degradation of insulin-like growth factor-binding protein-5 (IGFBP-5): regulation by IGFBP-5-(201–218).Am. J. Physiol. 1997; 273: E996-E1004
- O-Linked glycosylation in the mammary gland: changes that occur during malignancy.J. Mammary Gland Biol. Neoplasia. 2001; 6: 355-364
- Regulation and biological effect of endogenous insulin-like growth factor binding protein-5 in human osteoblastic cells.J. Clin. Endocrinol. Metab. 1993; 76: 1153-1159
- Heparin, heparan sulfate, and dermatan sulfate regulate formation of the insulin-like growth factor-I and insulin-like growth factor-binding protein complexes.J. Biol. Chem. 1994; 269: 20388-20393
- Binding of insulin-like growth factor (IGF)-binding protein-5 to smooth-muscle cell extracellular matrix is a major determinant of the cellular response to IGF-I.Mol. Biol. Cell. 1998; 9: 2383-2392
- Molecular distribution of IGF binding protein-5 in human serum.J. Clin. Endocrinol. Metab. 2002; 87: 271-276
- Structure and properties of the C-terminal domain of insulin-like growth factor-binding protein-1 isolated from human amniotic fluid.J. Biol. Chem. 2005; 280: 29812-29819
Published, MCP Papers in Press, May 11, 2007, DOI 10.1074/mcp.M700027-MCP200
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