Endocrinomic profile of neurointermediate lobe pituitary prohormone processing in PC1/3- and PC2-Null mice using SELDI-TOF mass spectrometry

Hardiman, Atira; Friedman, Theodore C.; Grunwald, William C.; Furuta, Machi; Zhu, Ziaorong; Steiner, Donald F.; Cool, David R.

doi:10.1677/jme.1.01812

Cited by 22 publications

(14 citation statements)

References 35 publications

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“…Comparative proteomics of the pituitary and hypothalamic extracts from WT and PCSK2-deficient mice have shown that several precursors to these peptides are either unprocessed or aberrantly processed. 11,28 The fact that the mutant mice show normal food intake despite impaired production of orexigenic peptides suggests that a balance between orexigenic and anorexigenic peptides may have been maintained. In this study, we measured the levels of the orexigenic gastric hormone ghrelin: they were comparable between WT and PCSK2-null mice.…”

Section: Discussionmentioning

confidence: 99%

Genetic deficiency for proprotein convertase subtilisin/kexin type 2 in mice is associated with decreased adiposity and protection from dietary fat-induced body weight gain

et al. 2010

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Background: Proprotein convertase subtilisin/xexin type 2 (PCSK2) is an endoproteinase responsible for proteolytic activation of a number of precursors to active neuropeptides and peptide hormones, known to influence glucose homeostasis, food intake and ultimately body mass. In this study, we examined the consequences of PCSK2 deficiency on these phenotypic traits. Study Design: Weight gain with age under diets of different fat contents was monitored. White adipose tissue (WAT) and muscle masses were evaluated. Plasma levels of triglycerides, leptin, ghrelin, insulin and proglucagon-derived peptides were measured as well as leptin and acetyl coenzyme-a carboxylase (ACCa) mRNA levels in adipose tissue. Results: Compared with their Pcsk2þ / þ littermates, Pcsk2 À/À mice weighed significantly less as weanlings and as adults. As adults, they carried noticeably less fat mass, with similar lean muscle mass: their plasma leptin level and adipose tissue leptin mRNA level were accordingly lower. PCSK2 deficiency did not affect food intake or the level of the orexigenic hormone ghrelin. However, PCSK2 deficiency resulted in decreased plasma triglycerides and reduced ACCa mRNA levels in WAT. Interestingly, unlike their Pcsk2 þ / þ littermates, Pcsk2 À/À were resistant to enhanced body weight gain when fed a high-fat diet. Consistent with a role of PCSK2 in body mass gain, diet-induced or genetically obese mice were found to contain significantly higher levels of PCSK2 mRNA in their brain and stomach than their lean counterparts. Conclusion: Collectively, these results suggest that PCSK2 contributes to increase in body mass through the various regulatory peptides generated through its action. It represents a potential target in the prevention and treatment of obesity.

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Section: Discussionmentioning

confidence: 99%

Genetic deficiency for proprotein convertase subtilisin/kexin type 2 in mice is associated with decreased adiposity and protection from dietary fat-induced body weight gain

et al. 2010

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“…In this well‐studied mouse model, selective roles of PC2 in determining the production of neuropeptide Y, somatostatin‐28, enkephalin, vaso‐active intestinal polypeptide (VIP), galanin and corticotropin releasing factor (CRF) were also demonstrated by RIA (Miller et al . 2003) and, recently, the first mass spectrometry‐based study of PC2‐null mice was performed, using a surface enhanced laser desorption ionization – time of flight (SELDI‐TOF) instrument (Hardiman et al . 2005).…”

Section: Discussionmentioning

confidence: 99%

Defective processing of neuropeptide precursors in Caenorhabditis elegans lacking proprotein convertase 2 (KPC‐2/EGL‐3): mutant analysis by mass spectrometry

Husson

Clynen

Baggerman

et al. 2006

Journal of Neurochemistry

128

154

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Biologically active peptides are synthesized as larger inactive proprotein peptide precursors which are processed by the concerted action of a cascade of enzymes. Among the proprotein convertases, PC2 is widely expressed in neuroendocrine tissues and has been proposed to be the major convertase involved in the biosynthesis of neuropeptides. In this study, we have examined the role of the Caenorhabditis elegans orthologue PC2/EGL-3 in the processing of proprotein peptide precursors. We recently isolated and identified 60 endogenous peptides in the nematode C. elegans by twodimensional nanoscale liquid chromatography -quadrupole time-of-flight tandem mass spectrometry. In the present study, we compare the peptide profile of different C. elegans strains, including PC2/EGL-3 mutants. For this purpose, we used an offline approach in which HPLC fractions are analysed by a matrix-assisted laser desorption ionisation -time of flight mass spectrometer. This differential peptidomic approach unambiguously provides evidence for the role of PC2/EGL-3 in the processing of FMRFamide-like peptide (FLP) precursors and neuropeptide-like protein (NLP) precursors in nematodes. Keywords: EGL-3, FMRFamide-like peptide gene, mass spectrometry, neuropeptide, neuropeptide-like protein gene, proprotein convertase. J. Neurochem. (2006Neurochem. ( ) 98, 1999Neurochem. ( -2012 Neuropeptides are derived from large proprotein peptide precursor genes, which encode for a single or multiple peptides. Several highly regulated and specific post-translational events are required for the formation of biologically active peptides from their larger inactive precursor proteins. A typical proprotein peptide precursor contains an amino terminal signal peptide, which is cleaved off upon entrance into the secretory pathway by a signal peptidase. Subsequently, proprotein convertases (PCs) cleave the remaining part of the precursor at specific cleavage sites. KR and RR constitute a large majority of all cleavage places, while RK and KK are found with much lower frequency (Veenstra 2000;Fricker 2005). In addition to these sites, some cleavages occur at sites containing pairs of basic amino acids separated by two, four, six or eight other residues. These sites were earlier described as 'monobasic' (Veenstra 2000). After cleavage from the proprotein peptide precursor, the peptide is further processed by a specific carboxypeptidase that removes the carboxyterminal basic amino acids. Next, the carboxyterminal glycine residue is transformed into an amide by a-amidating enzymes. This post-translational modification (conversion of a glycine into an amide) is a common feature of secretory peptides.In mammals, seven members of the kex2/subtilisin-like PCs have been described in detail (Rouille et al. 1995;Beinfeld 1998;Canaff et al. 1999). These are designated furin, PC1/3, PC2, PC4, PC5/6, PC7/8/LPC and PACE4 and they all share a characteristic overall similarity to the prototype member of the family, Kex2p, that was initially identified in yeast (Julius et a...

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“…1998; Allen et al . 2001), and provasopressin‐derived peptides showed no apparent change in level in PC2 KO mice tissue (Hardiman et al . 2005).…”

Section: Discussionmentioning

confidence: 99%

The role of prohormone convertase‐2 in hypothalamic neuropeptide processing: a quantitative neuropeptidomic study

Pan

Fei

Peng

et al. 2006

Journal of Neurochemistry

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Prohormone convertase (PC) 1/3 and 2 are involved in the generation of neuropeptides from their precursors. A quantitative peptidomic approach was used to explore the role PC2 plays in the processing of hypothalamic peptides. In this approach, extracts from mice lacking PC2 activity and from wildtype littermates were labeled with isotopic tags, combined, fractionated on a reverse phase HPLC column, and analyzed by electrospray ionization mass spectrometry. Altogether, 53 neuropeptides or other peptides derived from secretory pathway proteins were identified and sequenced using tandem mass spectrometry. These peptides arise from 21 distinct proteins: proenkephalin, proopiomelanocortin, prodynorphin, protachykinin A and B, procholecystokinin, promelanin-concentrating hormone, proneurotensin, proneuropeptide Y, provasopressin, pronociceptin/orphanin, prothyrotropinreleasing hormone, cocaine-and amphetamine-regulated transcript, chromogranin A and B, secretogranin II, prohormone convertase 1 and 2, propeptidyl-amidating monooxygenase, and proteins designated proSAAS and VGF. Approximately one third of the peptides found in wild-type mice were not detectable in PC2 knock-out mice, and another third were present at levels ranging from 25 to 75% of wild-type levels.Comparison of the cleavage sites suggests that sequences with a Trp, Tyr and/or Pro in the P1¢ or P2¢ position, or a basic residue in the P3 position, are preferentially cleaved by PC2 and not by other enzymes present in the secretory pathway. Keywords: carboxypeptidase E, neuropeptide biosynthesis, peptidase, prohormone convertase, proteomics, protease. Neuropeptides are generated from relatively large precursor molecules through limited proteolysis. The first step of proteolysis usually involves endoproteolytic cleavages at sites containing multiple basic amino acids such as KR, RR or RXXR (Rouille et al. 1995;Steiner 1998;Seidah et al. 1999;Zhou et al. 1999). After this step, peptide intermediates with basic C-terminal residues are further processed by a carboxypeptidase (Fricker 1988). In some cases, the resulting peptides are biologically active after the peptidase cleavages, while in other cases, additional post-translational modifications are required. These modifications include phosphorylation, acetylation, sulfation and C-terminal amidation (Mains et al. 1983).The endoproteolytic cleavages involved in the generation of neuroendocrine peptides are carried out by prohormone convertase 1 (PC1, also referred to as PC3 or PC1/3) and prohormone convertase 2 (PC2), which are members of a calcium-dependent, subtilisin-like serine protease family (Rouille et al. 1995;Seidah et al. 1999). These two enzymes are specifically expressed in neuroendocrine tissues and are enriched in peptide-containing secretory vesicles (Zheng et al. 1994;Rouille et al. 1995;Steiner 1998;Zhou et al. 1999). The enzymatic activities of both PC1/3 and PC2 have been extensively studied using in vitro and cell-line based approaches (Mathis and Lindberg 1992;Breslin et al. 1993...

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Endocrinomic profile of neurointermediate lobe pituitary prohormone processing in PC1/3- and PC2-Null mice using SELDI-TOF mass spectrometry

Cited by 22 publications

References 35 publications

Genetic deficiency for proprotein convertase subtilisin/kexin type 2 in mice is associated with decreased adiposity and protection from dietary fat-induced body weight gain

Genetic deficiency for proprotein convertase subtilisin/kexin type 2 in mice is associated with decreased adiposity and protection from dietary fat-induced body weight gain

Defective processing of neuropeptide precursors in Caenorhabditis elegans lacking proprotein convertase 2 (KPC‐2/EGL‐3): mutant analysis by mass spectrometry

The role of prohormone convertase‐2 in hypothalamic neuropeptide processing: a quantitative neuropeptidomic study

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