The heparin-binding domains of IGFBP-2 mediate its inhibitory effect on preadipocyte differentiation and fat development in male mice.

IGF-binding protein (IGFBP)-2 overexpression confers resistance to high-fat feeding and inhibits the differentiation of preadipocytes in vitro. However, whether administration of IGFBP-2 can regulate adipogenesis in vivo and the domains that mediate this response have not been defined. IGFBP-2 contains 2 heparin-binding domains (HBD), which are localized in the linker region (HBD1) and C-terminal region (HBD2) of IGFBP-2. To determine the relative importance of these domains, we used synthetic peptides as well as mutagenesis. Both HBD1 and HBD2 peptides inhibited preadipocyte differentiation, but the HBD2 peptide was more effective. Selective substitution of charged residues in the HBD1 or HBD2 regions attenuated the ability of the full-length protein to inhibit cell differentiation, but the HBD2 mutant had the greatest reduction. To determine their activities in vivo, pegylated forms of each peptide were administered to IGFBP-2(-/-) mice for 12 weeks. Magnetic resonance imaging scanning showed that only the HBD2 peptide significantly reduced (48 ± 9%, P < .05) gain in total fat mass. Both inguinal (32 ± 7%, P < .01) and visceral fat (44 ± 7%, P < .01) were significantly decreased by HBD2 whereas HBD1 reduced only visceral fat accumulation (24 ± 5%, P < .05). The HBD2 peptide was more effective peptide in reducing triglyceride content and serum adiponectin, but only the HBD2 peptide increased serum leptin. These findings demonstrate that the HBD2 domain of IGFBP-2 is the primary region that accounts for its ability to inhibit adipogenesis and that a peptide encompassing this region has activity that is comparable with native IGFBP-2.

[1]  M. Haluzík,et al.  Serum concentrations and tissue expression of components of insulin-like growth factor-axis in females with type 2 diabetes mellitus and obesity: The influence of very-low-calorie diet , 2012, Molecular and Cellular Endocrinology.

[2]  L. Maile,et al.  Insulin-Like Growth Factor (IGF) Binding Protein 2 Functions Coordinately with Receptor Protein Tyrosine Phosphatase β and the IGF-I Receptor To Regulate IGF-I-Stimulated Signaling , 2012, Molecular and Cellular Biology.

[3]  P. Poirier,et al.  Upregulation of Plasma Insulin‐Like Growth Factor Binding Protein 2 Levels After Biliopancreatic Diversion in Humans , 2012, Obesity.

[4]  L. Maile,et al.  Insulin‐like growth factor‐binding protein‐2 is required for osteoclast differentiation , 2012, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[5]  J. Ko,et al.  Influence of catch-up growth on IGFBP-2 levels and association between IGFBP-2 and cardiovascular risk factors in Korean children born SGA. , 2012, Endocrine journal.

[6]  Scott D Covey,et al.  Hepatic leptin signalling and subdiaphragmatic vagal efferents are not required for leptin-induced increases of plasma IGF binding protein-2 (IGFBP-2) in ob/ob mice , 2012, Diabetologia.

[7]  M. Bouxsein,et al.  The Heparin-binding Domain of IGFBP-2 Has Insulin-like Growth Factor Binding-independent Biologic Activity in the Growing Skeleton* , 2011, The Journal of Biological Chemistry.

[8]  V. Hwa,et al.  Acid-labile subunit (ALS) deficiency. , 2011, Best practice & research. Clinical endocrinology & metabolism.

[9]  T. Garland,et al.  Exercise, weight loss, and changes in body composition in mice: phenotypic relationships and genetic architecture. , 2011, Physiological genomics.

[10]  Bonadonna Riccardo,et al.  Adipocytes IGFBP‐2 Expression in Prepubertal Obese Children , 2010, Obesity.

[11]  F. Picard,et al.  Modulation of IGFBP2 mRNA expression in white adipose tissue upon aging and obesity. , 2010, Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme.

[12]  A. Hoeflich,et al.  Increased fat mass, decreased myofiber size, and a shift to glycolytic muscle metabolism in adolescent male transgenic mice overexpressing IGFBP-2. , 2010, American journal of physiology. Endocrinology and metabolism.

[13]  F. Hamdy,et al.  The relation between adiposity throughout the life course and variation in IGFs and IGFBPs: evidence from the ProtecT (Prostate testing for cancer and Treatment) study , 2010, Cancer Causes & Control.

[14]  A. Hokken-Koelega,et al.  Serum insulin-like growth factor-binding protein-2 levels and metabolic and cardiovascular risk factors in young adults and children born small for gestational age. , 2010, The Journal of clinical endocrinology and metabolism.

[15]  R. Wysocki,et al.  Antidiabetic effects of IGFBP2, a leptin-regulated gene. , 2010, Cell metabolism.

[16]  A. Pfeiffer,et al.  The role of insulin-like growth factor (IGF) binding protein-2 in the insulin-mediated decrease in IGF-I bioactivity. , 2009, The Journal of clinical endocrinology and metabolism.

[17]  S. Cummings,et al.  Serum Insulin‐Like Growth Factor‐1 Binding Proteins 1 and 2 and Mortality in Older Adults: The Health, Aging, and Body Composition Study , 2009, Journal of the American Geriatrics Society.

[18]  M. Kearney,et al.  IGF-dependent and IGF-independent actions of IGF-binding protein-1 and -2: implications for metabolic homeostasis , 2009, Trends in Endocrinology & Metabolism.

[19]  R. Baxter,et al.  Inhibition of adipocyte differentiation by insulin-like growth factor-binding protein-3. , 2009, American journal of physiology. Endocrinology and metabolism.

[20]  D. Clemmons,et al.  OR13,5 IGFBP-2 null mice develop obesity and insulin resistance with aging , 2008 .

[21]  D. Clemmons,et al.  p66shc negatively regulates insulin-like growth factor I signal transduction via inhibition of p52shc binding to Src homology 2 domain-containing protein tyrosine phosphatase substrate-1 leading to impaired growth factor receptor-bound protein-2 membrane recruitment. , 2008, Molecular endocrinology.

[22]  M. Bouxsein,et al.  Gender-specific changes in bone turnover and skeletal architecture in igfbp-2-null mice. , 2008, Endocrinology.

[23]  R. Norton,et al.  Cooperativity of the N- and C-terminal domains of insulin-like growth factor (IGF) binding protein 2 in IGF binding. , 2007, Biochemistry.

[24]  A. Juul,et al.  Effect of Weight Loss on Free Insulin‐Like Growth Factor‐I in Obese Women With Hyposomatotropism , 2007, Obesity.

[25]  K. Schmitz,et al.  Interactions between Insulin, Body Fat, and Insulin-Like Growth Factor Axis Proteins , 2007, Cancer Epidemiology Biomarkers & Prevention.

[26]  Antonio Vidal-Puig,et al.  IGF-Binding Protein-2 Protects Against the Development of Obesity and Insulin Resistance , 2007, Diabetes.

[27]  R. Norton,et al.  Structure, dynamics and heparin binding of the C-terminal domain of insulin-like growth factor-binding protein-2 (IGFBP-2). , 2006, Journal of molecular biology.

[28]  G. Davey Smith,et al.  Associations of adiposity from childhood into adulthood with insulin resistance and the insulin-like growth factor system: 65-year follow-up of the Boyd Orr Cohort. , 2006, The Journal of clinical endocrinology and metabolism.

[29]  S. Anderson,et al.  Insulin-like growth factor binding protein-2 (IGFBP-2) is a marker for the metabolic syndrome. , 2006, Experimental and clinical endocrinology & diabetes : official journal, German Society of Endocrinology [and] German Diabetes Association.

[30]  S. Chakrabarty,et al.  Insulin-like growth factor binding protein-2 stimulates proliferation and activates multiple cascades of the mitogen-activated protein kinase pathways in NIH-OVCAR3 human epithelial ovarian cancer cells , 2006, Cancer biology & therapy.

[31]  Seungho Wang,et al.  Inhibition of adipogenesis by RGD-dependent disintegrin. , 2005, Biochemical pharmacology.

[32]  A. Hoeflich,et al.  Insulin-like growth factor binding protein-2 binding to extracellular matrix plays a critical role in neuroblastoma cell proliferation, migration, and invasion. , 2005, Endocrinology.

[33]  A. Saltiel,et al.  Changes in integrin expression during adipocyte differentiation. , 2005, Cell metabolism.

[34]  G. Wright,et al.  Insulin-like growth factor binding protein 1 stimulates cell migration and binds to the a5I31 integrin by means of its Arg-Gly-Asp sequence , 2005 .

[35]  M. Elmlinger,et al.  Integrin-mediated action of insulin-like growth factor binding protein-2 in tumor cells. , 2004, Journal of molecular endocrinology.

[36]  D. Voskuil,et al.  Determinants of circulating insulin-like growth factor (IGF)-I and IGF binding proteins 1–3 in premenopausal women: physical activity and anthropometry (Netherlands) , 2001, Cancer Causes & Control.

[37]  J. Heinrich,et al.  Differential Impact of Simple Childhood Obesity on the Components of the Growth Hormone-Insulin-like Growth Factor (IGF)-IGF Binding Proteins Axis , 2004, Journal of pediatric endocrinology & metabolism : JPEM.

[38]  R. Baxter,et al.  Cellular actions of the insulin-like growth factor binding proteins. , 2002, Endocrine reviews.

[39]  S. Mohan,et al.  IGF-binding proteins are multifunctional and act via IGF-dependent and -independent mechanisms. , 2002, The Journal of endocrinology.

[40]  D. Clemmons,et al.  Use of mutagenesis to probe IGF-binding protein structure/function relationships. , 2001, Endocrine reviews.

[41]  R. Baxter,et al.  Mutagenesis of basic amino acids in the carboxyl-terminal region of insulin-like growth factor binding protein-5 affects acid-labile subunit binding. , 2001, Endocrinology.

[42]  D. Clemmons,et al.  Substitutions for Hydrophobic Amino Acids in the N-terminal Domains of IGFBP-3 and -5 Markedly Reduce IGF-I Binding and Alter Their Biologic Actions* , 2000, The Journal of Biological Chemistry.

[43]  G. Radetti,et al.  Growth hormone bioactivity, insulin-like growth factors (IGFs), and IGF binding proteins in obese children. , 1998, Metabolism: clinical and experimental.

[44]  W. Tamborlane,et al.  The metabolic syndrome and insulin-like growth factor I regulation in adolescent obesity. , 1998, The Journal of clinical endocrinology and metabolism.

[45]  J. Argente,et al.  Multiple endocrine abnormalities of the growth hormone and insulin-like growth factor axis in patients with anorexia nervosa: effect of short- and long-term weight recuperation. , 1997, The Journal of clinical endocrinology and metabolism.

[46]  Y. Song,et al.  Effect of obesity on total and free insulin-like growth factor (IGF)-1, and their relationship to IGF-binding protein (BP)-1, IGFBP-2, IGFBP-3, insulin, and growth hormone , 1997, International Journal of Obesity.

[47]  D. Clemmons,et al.  Identification of the Extracellular Matrix Binding Sites for Insulin-like Growth Factor-binding Protein 5* , 1996, The Journal of Biological Chemistry.

[48]  D. Clemmons,et al.  Insulin-like growth factor (IGF)-binding proteins inhibit the smooth muscle cell migration responses to IGF-I and IGF-II. , 1995, Endocrinology.

[49]  B. Moats-Staats,et al.  Expression of insulin-like growth factor-I (IGF-I) and IGF-binding proteins during adipogenesis. , 1994, Endocrinology.

[50]  E. Froesch,et al.  Regulation of insulin-like growth factor-I (IGF-I) and IGF-binding proteins by growth hormone in rat white adipose tissue. , 1993, Endocrinology.

[51]  D. Clemmons,et al.  Insulin-like growth factor binding protein 1 stimulates cell migration and binds to the alpha 5 beta 1 integrin by means of its Arg-Gly-Asp sequence. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[52]  M. Rechler,et al.  Transcriptional regulation of the rat IGFBP-1 and IGFBP-2 genes. , 1993, Growth regulation.

[53]  Y. Furukawa,et al.  A simple enzymatic quantitative analysis of triglycerides in tissues. , 1992, Journal of nutritional science and vitaminology.

[54]  N. Ling,et al.  Identification and molecular characterization of insulin-like growth factor binding proteins (IGFBP-1, -2, -3, -4, -5 and -6). , 1991, Progress in growth factor research.