Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling

R. M. Holmes; Z. Yi; E. De Filippis; R. Berria; S. Shahani; P. Sathyanarayana; V. Sherman; K. Fujiwara; C. Meyer; C. Christ-Roberts; H. Hwang; J. Finlayson; L. Q. Dong; L. J. Mandarino; M. Bajaj

doi:10.1007/s00125-011-2173-x

Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling

R. M. Holmes, Z. Yi, E. De Filippis, R. Berria, S. Shahani, P. Sathyanarayana, V. Sherman, K. Fujiwara, C. Meyer, C. Christ-Roberts, H. Hwang, J. Finlayson, L. Q. Dong, L. J. Mandarino, M. Bajaj

Endocrinology

Research output: Contribution to journal › Article › peer-review

33 Scopus citations

Abstract

Aims/hypothesis: The adiponectin signalling pathway is largely unknown, but recently the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1), has been shown to interact directly with adiponectin receptor (ADIPOR)1. APPL1 is present in C2C12 myoblasts and mouse skeletal muscle, but its presence in human skeletal muscle has not been investigated. Methods: Samples from type 2 diabetic, and lean and non-diabetic obese participants were analysed by: immunoprecipitation and western blot; HPLC-electrospray ionisation (ESI)-mass spectrometry (MS) analysis; peak area analysis by MS; HPLC-ESI-MS/MS/MS analysis; and RT-PCR analysis of APPL1 mRNA. Results: Immunoprecipitation and western blot indicated a band specific to APPL1. Tryptic digestion and HPLC-ESI-MS analysis of whole-muscle homogenate APPL1 unambiguously identified APPL1 with 56% sequence coverage. Peak area analysis by MS validated western blot results, showing APPL1 levels to be significantly increased in type 2 diabetic and obese as compared with lean participants. Targeted phosphopeptide analysis by HPLC-ESI-MS/MS/MS showed that APPL1 was phosphorylated specifically on Ser⁴⁰¹. APPL1 mRNA expression was significantly increased in obese and type 2 diabetic participants as compared with lean participants. After bariatric surgery in morbidly obese participants with subsequent weight loss, skeletal muscle APPL1 abundance was significantly reduced (p<0.05) in association with an increase in plasma adiponectin (p<0.01), increased levels of ADIPOR1 (p<0.05) and increased muscle AMP-activated protein kinase (AMPK) phosphorylation (p<0.05). Conclusions/interpretation: APPL1 abundance is significantly higher in type 2 diabetic muscle; APPL1 is phosphorylated in vivo on Ser ⁴⁰¹. Improvements in hyperglycaemia and hypoadiponectinaemia following weight loss are associated with reduced skeletal muscle APPL1, and increased plasma adiponectin levels and muscle AMPK phosphorylation.

Original language	English (US)
Pages (from-to)	2122-2131
Number of pages	10
Journal	Diabetologia
Volume	54
Issue number	8
DOIs	https://doi.org/10.1007/s00125-011-2173-x
State	Published - Aug 2011

Keywords

APPL1
Adiponectin
Obesity
Type 2 diabetes

ASJC Scopus subject areas

Internal Medicine
Endocrinology, Diabetes and Metabolism

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Holmes, R. M., Yi, Z., De Filippis, E., Berria, R., Shahani, S., Sathyanarayana, P., Sherman, V., Fujiwara, K., Meyer, C., Christ-Roberts, C., Hwang, H., Finlayson, J., Dong, L. Q., Mandarino, L. J., & Bajaj, M. (2011). Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling. Diabetologia, 54(8), 2122-2131. https://doi.org/10.1007/s00125-011-2173-x

Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling. / Holmes, R. M.; Yi, Z.; De Filippis, E. et al.
In: Diabetologia, Vol. 54, No. 8, 08.2011, p. 2122-2131.

Research output: Contribution to journal › Article › peer-review

Holmes, RM, Yi, Z, De Filippis, E, Berria, R, Shahani, S, Sathyanarayana, P, Sherman, V, Fujiwara, K, Meyer, C, Christ-Roberts, C, Hwang, H, Finlayson, J, Dong, LQ, Mandarino, LJ & Bajaj, M 2011, 'Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling', Diabetologia, vol. 54, no. 8, pp. 2122-2131. https://doi.org/10.1007/s00125-011-2173-x

Holmes RM, Yi Z, De Filippis E, Berria R, Shahani S, Sathyanarayana P et al. Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling. Diabetologia. 2011 Aug;54(8):2122-2131. doi: 10.1007/s00125-011-2173-x

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title = "Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling",

abstract = "Aims/hypothesis: The adiponectin signalling pathway is largely unknown, but recently the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1), has been shown to interact directly with adiponectin receptor (ADIPOR)1. APPL1 is present in C2C12 myoblasts and mouse skeletal muscle, but its presence in human skeletal muscle has not been investigated. Methods: Samples from type 2 diabetic, and lean and non-diabetic obese participants were analysed by: immunoprecipitation and western blot; HPLC-electrospray ionisation (ESI)-mass spectrometry (MS) analysis; peak area analysis by MS; HPLC-ESI-MS/MS/MS analysis; and RT-PCR analysis of APPL1 mRNA. Results: Immunoprecipitation and western blot indicated a band specific to APPL1. Tryptic digestion and HPLC-ESI-MS analysis of whole-muscle homogenate APPL1 unambiguously identified APPL1 with 56% sequence coverage. Peak area analysis by MS validated western blot results, showing APPL1 levels to be significantly increased in type 2 diabetic and obese as compared with lean participants. Targeted phosphopeptide analysis by HPLC-ESI-MS/MS/MS showed that APPL1 was phosphorylated specifically on Ser401. APPL1 mRNA expression was significantly increased in obese and type 2 diabetic participants as compared with lean participants. After bariatric surgery in morbidly obese participants with subsequent weight loss, skeletal muscle APPL1 abundance was significantly reduced (p<0.05) in association with an increase in plasma adiponectin (p<0.01), increased levels of ADIPOR1 (p<0.05) and increased muscle AMP-activated protein kinase (AMPK) phosphorylation (p<0.05). Conclusions/interpretation: APPL1 abundance is significantly higher in type 2 diabetic muscle; APPL1 is phosphorylated in vivo on Ser 401. Improvements in hyperglycaemia and hypoadiponectinaemia following weight loss are associated with reduced skeletal muscle APPL1, and increased plasma adiponectin levels and muscle AMPK phosphorylation.",

keywords = "APPL1, Adiponectin, Obesity, Type 2 diabetes",

author = "Holmes, {R. M.} and Z. Yi and {De Filippis}, E. and R. Berria and S. Shahani and P. Sathyanarayana and V. Sherman and K. Fujiwara and C. Meyer and C. Christ-Roberts and H. Hwang and J. Finlayson and Dong, {L. Q.} and Mandarino, {L. J.} and M. Bajaj",

note = "Funding Information: Acknowledgements This work was supported in part by grants from the American Diabetes Association 1-06-CR-03 (to M. Bajaj) and the Ron MacDonald Foundation at St Luke{\textquoteright}s Episcopal Hospital 08RDM008 (to M. Bajaj), as well as by a Marilyn Fishman Grant for Diabetes Research from the Endocrine Fellows Foundation (to S. Shahani) and grants NIH R01DK081750-01A1 (to Z. Yi) and NIH R01DK066483 (to L. J. Mandarino).",

year = "2011",

month = aug,

doi = "10.1007/s00125-011-2173-x",

language = "English (US)",

volume = "54",

pages = "2122--2131",

journal = "Diabetologia",

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TY - JOUR

T1 - Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes

T2 - Evidence for altered adiponectin signalling

AU - Holmes, R. M.

AU - Yi, Z.

AU - De Filippis, E.

AU - Berria, R.

AU - Shahani, S.

AU - Sathyanarayana, P.

AU - Sherman, V.

AU - Fujiwara, K.

AU - Meyer, C.

AU - Christ-Roberts, C.

AU - Hwang, H.

AU - Finlayson, J.

AU - Dong, L. Q.

AU - Mandarino, L. J.

AU - Bajaj, M.

N1 - Funding Information: Acknowledgements This work was supported in part by grants from the American Diabetes Association 1-06-CR-03 (to M. Bajaj) and the Ron MacDonald Foundation at St Luke’s Episcopal Hospital 08RDM008 (to M. Bajaj), as well as by a Marilyn Fishman Grant for Diabetes Research from the Endocrine Fellows Foundation (to S. Shahani) and grants NIH R01DK081750-01A1 (to Z. Yi) and NIH R01DK066483 (to L. J. Mandarino).

PY - 2011/8

Y1 - 2011/8

N2 - Aims/hypothesis: The adiponectin signalling pathway is largely unknown, but recently the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1), has been shown to interact directly with adiponectin receptor (ADIPOR)1. APPL1 is present in C2C12 myoblasts and mouse skeletal muscle, but its presence in human skeletal muscle has not been investigated. Methods: Samples from type 2 diabetic, and lean and non-diabetic obese participants were analysed by: immunoprecipitation and western blot; HPLC-electrospray ionisation (ESI)-mass spectrometry (MS) analysis; peak area analysis by MS; HPLC-ESI-MS/MS/MS analysis; and RT-PCR analysis of APPL1 mRNA. Results: Immunoprecipitation and western blot indicated a band specific to APPL1. Tryptic digestion and HPLC-ESI-MS analysis of whole-muscle homogenate APPL1 unambiguously identified APPL1 with 56% sequence coverage. Peak area analysis by MS validated western blot results, showing APPL1 levels to be significantly increased in type 2 diabetic and obese as compared with lean participants. Targeted phosphopeptide analysis by HPLC-ESI-MS/MS/MS showed that APPL1 was phosphorylated specifically on Ser401. APPL1 mRNA expression was significantly increased in obese and type 2 diabetic participants as compared with lean participants. After bariatric surgery in morbidly obese participants with subsequent weight loss, skeletal muscle APPL1 abundance was significantly reduced (p<0.05) in association with an increase in plasma adiponectin (p<0.01), increased levels of ADIPOR1 (p<0.05) and increased muscle AMP-activated protein kinase (AMPK) phosphorylation (p<0.05). Conclusions/interpretation: APPL1 abundance is significantly higher in type 2 diabetic muscle; APPL1 is phosphorylated in vivo on Ser 401. Improvements in hyperglycaemia and hypoadiponectinaemia following weight loss are associated with reduced skeletal muscle APPL1, and increased plasma adiponectin levels and muscle AMPK phosphorylation.

AB - Aims/hypothesis: The adiponectin signalling pathway is largely unknown, but recently the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1), has been shown to interact directly with adiponectin receptor (ADIPOR)1. APPL1 is present in C2C12 myoblasts and mouse skeletal muscle, but its presence in human skeletal muscle has not been investigated. Methods: Samples from type 2 diabetic, and lean and non-diabetic obese participants were analysed by: immunoprecipitation and western blot; HPLC-electrospray ionisation (ESI)-mass spectrometry (MS) analysis; peak area analysis by MS; HPLC-ESI-MS/MS/MS analysis; and RT-PCR analysis of APPL1 mRNA. Results: Immunoprecipitation and western blot indicated a band specific to APPL1. Tryptic digestion and HPLC-ESI-MS analysis of whole-muscle homogenate APPL1 unambiguously identified APPL1 with 56% sequence coverage. Peak area analysis by MS validated western blot results, showing APPL1 levels to be significantly increased in type 2 diabetic and obese as compared with lean participants. Targeted phosphopeptide analysis by HPLC-ESI-MS/MS/MS showed that APPL1 was phosphorylated specifically on Ser401. APPL1 mRNA expression was significantly increased in obese and type 2 diabetic participants as compared with lean participants. After bariatric surgery in morbidly obese participants with subsequent weight loss, skeletal muscle APPL1 abundance was significantly reduced (p<0.05) in association with an increase in plasma adiponectin (p<0.01), increased levels of ADIPOR1 (p<0.05) and increased muscle AMP-activated protein kinase (AMPK) phosphorylation (p<0.05). Conclusions/interpretation: APPL1 abundance is significantly higher in type 2 diabetic muscle; APPL1 is phosphorylated in vivo on Ser 401. Improvements in hyperglycaemia and hypoadiponectinaemia following weight loss are associated with reduced skeletal muscle APPL1, and increased plasma adiponectin levels and muscle AMPK phosphorylation.

KW - APPL1

KW - Adiponectin

KW - Obesity

KW - Type 2 diabetes

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Increased abundance of the adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif (APPL1) in patients with obesity and type 2 diabetes: Evidence for altered adiponectin signalling

Abstract

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