nach oben

Erschienen in:

01.11.2011 | Article

Global IRS-1 phosphorylation analysis in insulin resistance

verfasst von: P. Langlais, Z. Yi, J. Finlayson, M. Luo, R. Mapes, E. De Filippis, C. Meyer, E. Plummer, P. Tongchinsub, M. Mattern, L. J. Mandarino

Erschienen in: Diabetologia | Ausgabe 11/2011

Einloggen, um Zugang zu erhalten

Abstract

Aims/hypothesis

IRS-1 serine phosphorylation is often elevated in insulin resistance models, but confirmation in vivo in humans is lacking. We therefore analysed IRS-1 phosphorylation in human muscle in vivo.

Methods

We used HPLC-electrospray ionisation (ESI)-MS/MS to quantify IRS-1 phosphorylation basally and after insulin infusion in vastus lateralis muscle from lean healthy, obese non-diabetic and type 2 diabetic volunteers.

Results

Basal Ser323 phosphorylation was increased in type 2 diabetic patients (2.1 ± 0.43, p ≤ 0.05, fold change vs lean controls). Thr495 phosphorylation was decreased in type 2 diabetic patients (p ≤ 0.05). Insulin increased IRS-1 phosphorylation at Ser527 (1.4 ± 0.17, p ≤ 0.01, fold change, 60 min after insulin infusion vs basal) and Ser531 (1.3 ± 0.16, p ≤ 0.01, fold change, 60 min after insulin infusion vs basal) in the lean controls and suppressed phosphorylation at Ser348 (0.56 ± 0.11, p ≤ 0.01, fold change, 240 min after insulin infusion vs basal), Thr446 (0.64 ± 0.16, p ≤ 0.05, fold change, 60 min after insulin infusion vs basal), Ser1100 (0.77 ± 0.22, p ≤ 0.05, fold change, 240 min after insulin infusion vs basal) and Ser1142 (1.3 ± 0.2, p ≤ 0.05, fold change, 60 min after insulin infusion vs basal).

Conclusions/interpretation

We conclude that, unlike some aspects of insulin signalling, the ability of insulin to increase or suppress certain IRS-1 phosphorylation sites is intact in insulin resistance. However, some IRS-1 phosphorylation sites do not respond to insulin, whereas other Ser/Thr phosphorylation sites are either increased or decreased in insulin resistance.

Nur mit Berechtigung zugänglich

Gual P, Le Marchand-Brustel Y, Tanti JF (2005) Positive and negative regulation of insulin signaling through IRS-1 phosphorylation. Biochimie 87:99–109PubMedCrossRef

Zick Y (2003) Role of Ser/Thr kinases in the uncoupling of insulin signaling. Int J Obes Relat Metab Disord 27(Suppl 3):S56–S60PubMedCrossRef

Haruta T, Uno T, Kawahara J et al (2000) A rapamycin-sensitive pathway down-regulates insulin signaling via phosphorylation and proteasomal degradation of insulin receptor substrate-1. Mol Endocrinol 14:783–794PubMedCrossRef

Takano A, Usui I, Haruta T et al (2001) Mammalian target of rapamycin pathway regulates insulin signaling via subcellular redistribution of insulin receptor substrate 1 and integrates nutritional signals and metabolic signals of insulin. Mol Cell Biol 21:5050–5062PubMedCrossRef

Sun XJ, Goldberg JL, Qiao LY, Mitchell JJ (1999) Insulin-induced insulin receptor substrate-1 degradation is mediated by the proteasome degradation pathway. Diabetes 48:1359–1364PubMedCrossRef

Pederson TM, Kramer DL, Rondinone CM (2001) Serine/threonine phosphorylation of IRS-1 triggers its degradation: possible regulation by tyrosine phosphorylation. Diabetes 50:24–31PubMedCrossRef

Shah OJ, Wang Z, Hunter T (2004) Inappropriate activation of the TSC/Rheb/mTOR/S6K cassette induces IRS1/2 depletion, insulin resistance, and cell survival deficiencies. Curr Biol 14:1650–1656PubMedCrossRef

Paz K, Hemi R, LeRoith D et al (1997) A molecular basis for insulin resistance. Elevated serine/threonine phosphorylation of IRS-1 and IRS-2 inhibits their binding to the juxtamembrane region of the insulin receptor and impairs their ability to undergo insulin-induced tyrosine phosphorylation. J Biol Chem 272:29911–29918PubMedCrossRef

Liu YF, Paz K, Herschkovitz A et al (2001) Insulin stimulates PKCzeta -mediated phosphorylation of insulin receptor substrate-1 (IRS-1). A self-attenuated mechanism to negatively regulate the function of IRS proteins. J Biol Chem 276:14459–14465PubMed

10.

Ravichandran LV, Esposito DL, Chen J, Quon MJ (2001) Protein kinase C-zeta phosphorylates insulin receptor substrate-1 and impairs its ability to activate phosphatidylinositol 3-kinase in response to insulin. J Biol Chem 276:3543–3549PubMedCrossRef

11.

Moeschel K, Beck A, Weigert C et al (2004) Protein kinase C-zeta-induced phosphorylation of Ser318 in insulin receptor substrate-1 (IRS-1) attenuates the interaction with the insulin receptor and the tyrosine phosphorylation of IRS-1. J Biol Chem 279:25157–25163PubMedCrossRef

12.

Aguirre V, Uchida T, Yenush L, Davis R, White MF (2000) The c-Jun NH(2)-terminal kinase promotes insulin resistance during association with insulin receptor substrate-1 and phosphorylation of Ser(307). J Biol Chem 275:9047–9054PubMedCrossRef

13.

Liu YF, Herschkovitz A, Boura-Halfon S et al (2004) Serine phosphorylation proximal to its phosphotyrosine binding domain inhibits insulin receptor substrate 1 function and promotes insulin resistance. Mol Cell Biol 24:9668–9681PubMedCrossRef

14.

Aguirre V, Werner ED, Giraud J, Lee YH, Shoelson SE, White MF (2002) Phosphorylation of Ser307 in insulin receptor substrate-1 blocks interactions with the insulin receptor and inhibits insulin action. J Biol Chem 277:1531–1537PubMedCrossRef

15.

Mothe I, Van Obberghen E (1996) Phosphorylation of insulin receptor substrate-1 on multiple serine residues, 612, 632, 662, and 731, modulates insulin action. J Biol Chem 271:11222–11227PubMedCrossRef

16.

Li J, DeFea K, Roth RA (1999) Modulation of insulin receptor substrate-1 tyrosine phosphorylation by an Akt/phosphatidylinositol 3-kinase pathway. J Biol Chem 274:9351–9356PubMedCrossRef

17.

Paz K, Liu YF, Shorer H et al (1999) Phosphorylation of insulin receptor substrate-1 (IRS-1) by protein kinase B positively regulates IRS-1 function. J Biol Chem 274:28816–28822PubMedCrossRef

18.

Gual P, Gremeaux T, Gonzalez T, Le Marchand-Brustel Y, Tanti JF (2003) MAP kinases and mTOR mediate insulin-induced phosphorylation of insulin receptor substrate-1 on serine residues 307, 612 and 632. Diabetologia 46:1532–1542PubMedCrossRef

19.

Rui L, Aguirre V, Kim JK et al (2001) Insulin/IGF-1 and TNF-alpha stimulate phosphorylation of IRS-1 at inhibitory Ser307 via distinct pathways. J Clin Invest 107:181–189PubMedCrossRef

20.

Copps KD, Hancer NJ, Opare-Ado L, Qiu W, Walsh C, White MF (2010) Irs1 serine 307 promotes insulin sensitivity in mice. Cell Metab 11:84–92PubMedCrossRef

21.

Werner ED, Lee J, Hansen L, Yuan M, Shoelson SE (2004) Insulin resistance due to phosphorylation of insulin receptor substrate-1 at serine 302. J Biol Chem 279:35298–35305PubMedCrossRef

22.

Yi Z, Langlais P, De Filippis EA et al (2007) Global assessment of regulation of phosphorylation of insulin receptor substrate-1 by insulin in vivo in human muscle. Diabetes 56:1508–1516PubMedCrossRef

23.

Langlais P, Mandarino LJ, Yi Z (2010) Label-free relative quantification of co-eluting isobaric phosphopeptides of insulin receptor substrate-1 by HPLC-ESI-MS/MS. J Am Soc Mass Spectrom 21:1490–1499PubMedCrossRef

24.

Cusi K, Maezono K, Osman A et al (2000) Insulin resistance differentially affects the PI 3-kinase- and MAP kinase-mediated signaling in human muscle. J Clin Invest 105:311–320PubMedCrossRef

25.

DeFronzo RA, Tobin JD, Andres R (1979) Glucose clamp technique: a method for quantifying insulin secretion and resistance. Am J Physiol 237:E214–E223PubMed

26.

Lefort N, Glancy B, Bowen B et al (2010) Increased reactive oxygen species production and lower abundance of complex I subunits and carnitine palmitoyltransferase 1B protein despite normal mitochondrial respiration in insulin-resistant human skeletal muscle. Diabetes 59:2444–2452PubMedCrossRef

27.

Sriwijitkamol A, Christ-Roberts C, Berria R et al (2006) Reduced skeletal muscle inhibitor of kappaB beta content is associated with insulin resistance in subjects with type 2 diabetes: reversal by exercise training. Diabetes 55:760–767PubMedCrossRef

28.

Debodo RC, Steele R, Altszuler N, Dunn A, Bishop JS (1963) On the hormonal regulation of carbohydrate metabolism; studies with C14 glucose. Recent Prog Horm Res 19:445–488PubMed

29.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMed

30.

Griffin ME, Marcucci MJ, Cline GW et al (1999) Free fatty acid-induced insulin resistance is associated with activation of protein kinase C theta and alterations in the insulin signaling cascade. Diabetes 48:1270–1274PubMedCrossRef

31.

Lee YH, Giraud J, Davis RJ, White MF (2003) c-Jun N-terminal kinase (JNK) mediates feedback inhibition of the insulin signaling cascade. J Biol Chem 278:2896–2902PubMedCrossRef

32.

Hirosumi J, Tuncman G, Chang L et al (2002) A central role for JNK in obesity and insulin resistance. Nature 420:333–336PubMedCrossRef

33.

Gao Z, Zhang X, Zuberi A et al (2004) Inhibition of insulin sensitivity by free fatty acids requires activation of multiple serine kinases in 3T3-L1 adipocytes. Mol Endocrinol 18:2024–2034PubMedCrossRef

34.

Nguyen MT, Satoh H, Favelyukis S et al (2005) JNK and tumor necrosis factor-alpha mediate free fatty acid-induced insulin resistance in 3T3-L1 adipocytes. J Biol Chem 280:35361–35371PubMedCrossRef

35.

Yuan M, Konstantopoulos N, Lee J et al (2001) Reversal of obesity- and diet-induced insulin resistance with salicylates or targeted disruption of Ikkbeta. Science 293:1673–1677PubMedCrossRef

36.

Perseghin G, Petersen K, Shulman GI (2003) Cellular mechanism of insulin resistance: potential links with inflammation. Int J Obes Relat Metab Disord 27(Suppl 3):S6–S11PubMedCrossRef

37.

Gao Z, Hwang D, Bataille F et al (2002) Serine phosphorylation of insulin receptor substrate 1 by inhibitor kappa B kinase complex. J Biol Chem 277:48115–48121PubMedCrossRef

38.

Um SH, Frigerio F, Watanabe M et al (2004) Absence of S6K1 protects against age- and diet-induced obesity while enhancing insulin sensitivity. Nature 431:200–205PubMedCrossRef

39.

Pende M, Kozma SC, Jaquet M et al (2000) Hypoinsulinaemia, glucose intolerance and diminished beta-cell size in S6K1-deficient mice. Nature 408:994–997PubMedCrossRef

40.

Tremblay F, Krebs M, Dombrowski L et al (2005) Overactivation of S6 kinase 1 as a cause of human insulin resistance during increased amino acid availability. Diabetes 54:2674–2684PubMedCrossRef

41.

Greene MW, Morrice N, Garofalo RS, Roth RA (2004) Modulation of human insulin receptor substrate-1 tyrosine phosphorylation by protein kinase Cdelta. Biochem J 378:105–116PubMedCrossRef

42.

Weigert C, Hennige AM, Brischmann T et al (2005) The phosphorylation of Ser318 of insulin receptor substrate 1 is not per se inhibitory in skeletal muscle cells but is necessary to trigger the attenuation of the insulin-stimulated signal. J Biol Chem 280:37393–37399PubMedCrossRef

43.

Weigert C, Kron M, Kalbacher H et al (2008) Interplay and effects of temporal changes in the phosphorylation state of serine-302, -307, and -318 of insulin receptor substrate-1 on insulin action in skeletal muscle cells. Mol Endocrinol 22:2729–2740PubMedCrossRef

44.

Zick Y (2005) Ser/Thr phosphorylation of IRS proteins: a molecular basis for insulin resistance. Sci STKE 2005:pe4PubMedCrossRef

45.

Bouzakri K, Roques M, Gual P et al (2003) Reduced activation of phosphatidylinositol-3 kinase and increased serine 636 phosphorylation of insulin receptor substrate-1 in primary culture of skeletal muscle cells from patients with type 2 diabetes. Diabetes 52:1319–1325PubMedCrossRef

46.

Tzatsos A, Kandror KV (2006) Nutrients suppress phosphatidylinositol 3-kinase/Akt signaling via raptor-dependent mTOR-mediated insulin receptor substrate 1 phosphorylation. Mol Cell Biol 26:63–76PubMedCrossRef

47.

Langlais P, Yi Z, Mandarino LJ (2011) The identification of raptor as a substrate for p44/42 MAPK. Endocrinology 152:1264–1273PubMedCrossRef

48.

Giraud J, Haas M, Feener EP et al (2007) Phosphorylation of Irs1 at SER-522 inhibits insulin signaling. Mol Endocrinol 21:2294–2302PubMedCrossRef

Titel: Global IRS-1 phosphorylation analysis in insulin resistance
verfasst von: P. Langlais
Z. Yi
J. Finlayson
M. Luo
R. Mapes
E. De Filippis
C. Meyer
E. Plummer
P. Tongchinsub
M. Mattern
L. J. Mandarino
Publikationsdatum: 01.11.2011
Verlag: Springer-Verlag
Erschienen in: Diabetologia / Ausgabe 11/2011
Print ISSN: 0012-186X
Elektronische ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-011-2271-9

Leitlinien kompakt für die Innere Medizin

Mit medbee Pocketcards sicher entscheiden.

^{Seit 2022 gehört die medbee GmbH zum Springer Medizin Verlag}

Kostenlos registrieren

Update Innere Medizin

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.

Newsletter bestellen

Live-Webinar "Urologie und Sexualmedizin in der Praxis"

Springer Medizin

Global IRS-1 phosphorylation analysis in insulin resistance