Biological Processing of the Cocaine and Amphetamine-regulated Transcript Precursors by Prohormone Convertases, PC2 and PC1/3

Dey, Arunangsu; Xhu, Xiaorong; Carroll, Raymond J.; Turck, C. W.; Stein, Jeffrey A.; Steiner, Donald F.

doi:10.1074/jbc.m212128200

Cited by 76 publications

(52 citation statements)

References 57 publications

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“…To measure individual band intensity, background was subtracted for each protein band followed by normalization against the control protein (tubulin). (E) a-e, isolated adult human islets from four independent batches (two [HI-1 and HI-2] from low BMI [26][27] and two [HI-3 and HI-4] from high BMI [40][41][42][43][44][45][46][47][48][49][50] cadaveric donors) with high purity (80-90% for low BMI islets) and low purity (35-40% for high BMI islets) were cultured and equal amounts (50 μg) of human islet lysates or INS-1 CE were fractionated in SDS-PAGE and the transferred peptides were incubated with specific primary antibodies as shown. *p < 0.05; NS, not significant.…”

Section: Resultsmentioning

confidence: 99%

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GSK-3 inactivation or depletion promotes β-cell replication via down regulation of the CDK inhibitor, p27 (Kip1)

Stein¹,

Milewski²,

Hara³

et al. 2011

Islets

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Section: Resultsmentioning

confidence: 99%

“…INS-1 cells and human islets were brought into ice-cold immunoprecipitation (IP) buffer containing freshly added protease inhibitor cocktails 50 and phosphatase inhibitor cocktails A and B (Santa Cruz Biotechnology Inc.). Cells/islets were sonicated and supernatants were collected after microcentrifugation.…”

mentioning

confidence: 99%

GSK-3 inactivation or depletion promotes β-cell replication via down regulation of the CDK inhibitor, p27 (Kip1)

Stein¹,

Milewski²,

Hara³

et al. 2011

Islets

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“…Metabolic labeling, immunoprecipitation, and Western blotting were performed as described before (35). Aprotinin (5 g/ml) was added in chase medium containing 10ϫ excess cold methionine following metabolic pulse labeling.…”

Section: Methodsmentioning

confidence: 99%

Altered Proglucagon Processing in an α-Cell Line Derived from Prohormone Convertase 2 Null Mouse Islets

Webb

Dey²,

Wang³

et al. 2004

Journal of Biological Chemistry

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The endoproteolytic processing of proproteins in the secretory pathway depends on the expression of selected members of a family of subtilisin-like endoproteases known as the prohormone convertases (PCs). The main PC family members expressed in mammalian neuroendocrine cells are PC2 and PC1/3. The differential processing of proglucagon in pancreatic ␣-cells and intestinal L cells leads to production of distinct hormonal products with opposing physiological effects from the same precursor. Here we describe the establishment and characterization of a novel ␣-cell line (␣TC-⌬PC2) derived from PC2 homozygous null animals. The ␣TC-⌬PC2 cells are shown to be similar to the well characterized ␣TC1-6 cell line in both morphology and overall gene expression. However, the absence of PC2 activity in ␣TC-⌬PC2 leads to a complete block in the production of mature glucagon. Surprisingly, ␣TC-⌬PC2 cells are able to efficiently cleave the interdomain site in proglucagon (KR 70 -71). Further analysis reveals that ␣TC-⌬PC2 cells, unlike ␣TC1-6 cells, express low levels of PC1/3 that lead to the generation of glicentin as well as low amounts of oxyntomodulin, GLP-1, truncated GLP-1, and N-terminally extended GLP-2. We conclude that ␣TC-⌬PC2 cells provide additional evidence for PC2 as the major convertase in ␣-cells leading to mature glucagon production and provide a robust model for further analysis of the mechanisms of proprotein processing by the prohormone convertases.

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“…In the hypothalamus, these enzymes displayed an extensive overlapping pattern of expression. Both convertases are effectively targeted to the regulated secretory pathway, where they process many neuropeptide precursors in the brain and prohormones in the pituitary, including pro-TRH (7,24), POMC (9), prosomatostatin (25), provasopressin (26), pro-neuropeptide Y (proNPY) (27), proneurotensin (28), proenkephalin (29), and pro-cocaine amphetamine-regulated transcript (proCART) (30). The critical role of PC1 and PC2 in prohormone processing is underscored by studies of animals that lack the genes encoding PC1 (31) and PC2 (32), as well as 7B2 (33,34), a neuropeptide essential for the maturation of PC2.…”

Section: Introductionmentioning

confidence: 99%

Regulation of hypothalamic prohormone convertases 1 and 2 and effects on processing of prothyrotropin-releasing hormone

Sanchez¹,

Goldstein²,

Stuart³

et al. 2004

J. Clin. Invest.

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Regulation of energy balance by leptin involves regulation of several neuropeptides, including thyrotropinreleasing hormone (TRH).Synthesized from a larger inactive precursor, its maturation requires proteolytic cleavage by prohormone convertases 1 and 2 (PC1 and PC2). Since this maturation in response to leptin requires prohormone processing, we hypothesized that leptin might regulate hypothalamic PC1 and PC2 expression, ultimately leading to coordinated processing of prohormones into mature peptides. Using hypothalamic neurons, we found that leptin stimulated PC1 and PC2 mRNA and protein expression and also increased PC1 and PC2 promoter activities in transfected 293T cells. Starvation of rats, leading to low serum leptin levels, decreased PC1 and PC2 gene and protein expression in the paraventricular nucleus (PVN) of the hypothalamus. Exogenous administration of leptin to fasted animals restored PC1 levels in the median eminence (ME) and the PVN to approximately the level found in fed control animals. Consistent with this regulation of PCs in the PVN, concentrations of TRH in the PVN and ME were substantially reduced in the fasted animals relative to the fed animals, and leptin reversed this decrease. Further analysis showed that proteolytic cleavage of pro-thyrotropin-releasing hormone (proTRH) at known PC cleavage sites was reduced by fasting and increased in animals given leptin. Combined, these findings suggest that leptin-dependent stimulation of hypothalamic TRH expression involves both activation of trh transcription and stimulation of PC1 and PC2 expression, which lead to enhanced processing of proTRH into mature TRH.

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Biological Processing of the Cocaine and Amphetamine-regulated Transcript Precursors by Prohormone Convertases, PC2 and PC1/3

Cited by 76 publications

References 57 publications

GSK-3 inactivation or depletion promotes β-cell replication via down regulation of the CDK inhibitor, p27 (Kip1)

GSK-3 inactivation or depletion promotes β-cell replication via down regulation of the CDK inhibitor, p27 (Kip1)

Altered Proglucagon Processing in an α-Cell Line Derived from Prohormone Convertase 2 Null Mouse Islets

Regulation of hypothalamic prohormone convertases 1 and 2 and effects on processing of prothyrotropin-releasing hormone

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