{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T11:26:26Z","timestamp":1768994786333,"version":"3.49.0"},"reference-count":52,"publisher":"The Endocrine Society","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2013,3,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We previously demonstrated that hypoxia and leucine deprivation cause hyperphosphorylation of IGF-binding protein-1 (IGFBP-1) at discrete sites that markedly enhanced IGF-I affinity and inhibited IGF-I-stimulated cell growth. In this study we investigated the functional role of these phosphorylation sites using mutagenesis. We created three IGFBP-1 mutants in which individual serine (S119\/S169\/S98) residues were substituted with alanine and S101A was recreated for comparison. The wild-type (WT) and mutant IGFBP-1 were expressed in Chinese hamster ovary cells and IGFBP-1 in cell media was isolated using isoelectric-focusing-free-flow electrophoresis. BIACore analysis indicated that the changes in IGF-I affinity for S98A and S169A were moderate, whereas S119A greatly reduced the affinity of IGFBP-1 for IGF-I (100-fold, P &amp;lt; .0001). Similar results were obtained with S101A. The IGF-I affinity changes of the mutants were reflected in their ability to inhibit IGF-I-induced receptor autophosphorylation. Employing receptor-stimulation assay using IGF-IR-overexpressing P6 cells, we found that WT-IGFBP-1 inhibited IGF-IR\u03b2 autophosphorylation (\u223c2-fold, P &amp;lt; .001), possibly attributable to sequestration of IGF-I. Relative to WT, S98A and S169A mutants did not inhibit receptor autophosphorylation. S119A, on the other hand, greatly stimulated the receptor (2.3-fold, P &amp;lt; .05). The data with S101A matched S119A. In summary, we show that phosphorylation at S98 and S169 resulted in milder changes in IGF-I action; nonetheless most dramatic inhibitory effects on the biological activity of IGF-I were due to IGFBP-1 phosphorylation at S119. Our results provide novel demonstration that IGFBP-1 phosphorylation at S119 can enhance affinity for IGF-I possibly through stabilization of the IGF-IGFBP-1 complex. These data also propose that the synergistic interaction of distinct phosphorylation sites may be important in eliciting more pronounced effects on IGF-I affinity that needs further investigation.<\/jats:p>","DOI":"10.1210\/en.2012-1962","type":"journal-article","created":{"date-parts":[[2013,1,26]],"date-time":"2013-01-26T03:30:29Z","timestamp":1359171029000},"page":"1130-1143","source":"Crossref","is-referenced-by-count":27,"title":["Phosphorylation of IGFBP-1 at Discrete Sites Elicits Variable Effects on IGF-I Receptor Autophosphorylation"],"prefix":"10.1210","volume":"154","author":[{"given":"Majida","family":"Abu Shehab","sequence":"first","affiliation":[{"name":"Children's Health Research Institute (M.A.S., C.I., M.B.G.), Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cristiana","family":"Iosef","sequence":"first","affiliation":[{"name":"Children's Health Research Institute (M.A.S., C.I., M.B.G.), Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert","family":"Wildgruber","sequence":"first","affiliation":[{"name":"Free Flow Electrophoresis Service GmbH (R.W.), Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Girish","family":"Sardana","sequence":"first","affiliation":[{"name":"Ontario Cancer Biomarker Network MaRS (G.S.), Toronto, Ontario M5G 1L7 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Madhulika B.","family":"Gupta","sequence":"first","affiliation":[{"name":"Department of Pediatrics (M.B.G.) and Biochemistry (M.B.G.), The Western University, London, Ontario N6C 2V5 Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"80","reference":[{"key":"2020071612044779200_B1","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/0955-2235(95)00013-5","article-title":"Role of insulin-like growth factor binding proteins in the control of IGF actions","volume":"6","author":"Clemmons","year":"1995","journal-title":"Prog Growth Factor Res"},{"key":"2020071612044779200_B2","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/S0303-7207(98)00024-0","article-title":"Role of insulin-like growth factor binding proteins in controlling IGF actions","volume":"140","author":"Clemmons","year":"1998","journal-title":"Mol Cell Endocrinol"},{"key":"2020071612044779200_B3","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/0955-2235(95)00015-1","article-title":"Cell migration: interactions among integrins, IGFs and IGFBPs","volume":"6","author":"Jones","year":"1995","journal-title":"Prog Growth Factor Res"},{"key":"2020071612044779200_B4","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1210\/er.2001-0033","article-title":"Cellular actions of the insulin-like growth factor binding proteins","volume":"23","author":"Firth","year":"2002","journal-title":"Endocr Rev"},{"key":"2020071612044779200_B5","first-page":"801","article-title":"Insulin-like growth factor-binding proteins in serum and other biological fluids: regulation and functions","volume":"18","author":"Rajaram","year":"1997","journal-title":"Endocr Rev"},{"key":"2020071612044779200_B6","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/1471-2199-5-15","article-title":"Glycogen synthase kinase-3 regulates IGFBP-1 gene transcription through the thymine-rich insulin response element","volume":"5","author":"Finlay","year":"2004","journal-title":"BMC Mol Biol"},{"key":"2020071612044779200_B7","doi-asserted-by":"crossref","first-page":"10188","DOI":"10.1073\/pnas.95.17.10188","article-title":"Hypoxia stimulates insulin-like growth factor binding protein 1 (IGFBP-1) gene expression in HepG2 cells: a possible model for IGFBP-1 expression in fetal hypoxia","volume":"95","author":"Tazuke","year":"1998","journal-title":"Proc Natl Acad Sci USA"},{"key":"2020071612044779200_B8","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1093\/oxfordjournals.humrep.a137426","article-title":"Insulin-like growth factor binding protein-1 inhibits the DNA amplification induced by insulin-like growth factor I in human granulosa-luteal cells","volume":"6","author":"Angervo","year":"1991","journal-title":"Hum Reprod"},{"key":"2020071612044779200_B9","doi-asserted-by":"crossref","first-page":"14203","DOI":"10.1016\/S0021-9258(18)68206-7","article-title":"Purification of a 31,000-dalton insulin-like growth factor binding protein from human amniotic fluid. 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