Prohormone convertase 1/3 is essential for processing of the glucose-dependent insulinotropic polypeptide precursor

Randi Ugleholdt; Marie-Louise H Poulsen; Peter J Holst; Jean-Claude Irminger; Cathrine Orskov; Jens Pedersen; Mette M Rosenkilde; Xiaorong Zhu; Donald F Steiner; Jens Juul Holst

doi:10.1074/jbc.M601203200

Prohormone convertase 1/3 is essential for processing of the glucose-dependent insulinotropic polypeptide precursor

Bidragets oversatte titel: Prohormone convertase 1 / 3 af afgørende betydning for behandlingen af glukose-insulinotropt polypeptid forløber

Randi Ugleholdt, Marie-Louise H Poulsen, Peter J Holst, Jean-Claude Irminger, Cathrine Orskov, Jens Pedersen, Mette M Rosenkilde, Xiaorong Zhu, Donald F Steiner, Jens Juul Holst

71 Citationer (Scopus)

Abstract

Fysiologi inkretinhormonerne, glukagon-lignende peptid 1 (GLP-1) og glucose-insulinotropt polypeptid (GIP), og deres rolle i type 2-diabetes i øjeblikket tiltrækker stor interesse. Vi har for nylig rapporteret en afgørende rolle for prohormone convertase (PC) 1 / 3 i spaltningen af intestinal proglucagon, hvilket resulterer i dannelsen af GLP-1, som vist i PC1/3-deficient mus. Imidlertid kun lidt om den endoproteolytic behandlingen af GIP forløber. Dette studie undersøger behandling af proGIP i PC1 / 3 og PC2 null-mus og i cellelinier ved hjælp af adenovirus-medieret overekspression. Støtte til en rolle for PC1 / 3 i proGIP behandling, fandt vi co-lokalisering af GIP og PC1 / 3, men ikke PC2 i tarm sektioner ved immunhistokemi, og analyse af intestinale ekstrakter fra PC1/3-deficient dyr vist alvorligt svækket behandling til GIP, der henviser til behandling for at GIP var uændret i PC2-deficiente mus. Derfor overekspression af preproGIP i neuroendokrine ATT-20 cellelinie, der udtrykker et højt niveau af endogen PC1 / 3 og ubetydelige niveauer af PC2 resulteret i produktion af GIP. Lignende resultater blev opnået efter co-ekspression af preproGIP og PC1 / 3 i GH4 celler, der udtrykker noget PC2 og kun lave niveauer af PC1 / 3. Desuden viser undersøgelser i GH4 celler og alfa-TC1.9 cellelinie, der udtrykker PC2 men ikke PC1 / 3, at PC2 kan mægle forarbejdning til GIP, men også til andre fragmenter ikke fundet i tarm-ekstrakter. Tilsammen vores data viser, at PC1 / 3 er vigtig og tilstrækkelig til produktion af den intestinale inkretin hormonet GIP, mens PC2 selv i stand til at kløve proGIP, deltager ikke i tarm proGIP forarbejdning, og er ikke fundet i tarm GIP-udtrykkende celler .
Udgivelsesdato: 2006-Apr-21

Bidragets oversatte titel	Prohormone convertase 1 / 3 af afgørende betydning for behandlingen af glukose-insulinotropt polypeptid forløber
Originalsprog	Engelsk
Tidsskrift	Journal of Biological Chemistry
Vol/bind	281
Udgave nummer	16
Sider (fra-til)	11050-7
Antal sider	7
ISSN	0021-9258
DOI	https://doi.org/10.1074/jbc.M601203200
Status	Udgivet - 2006

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@article{a1aa5f409d2d11debc73000ea68e967b,

title = "Prohormone convertase 1/3 is essential for processing of the glucose-dependent insulinotropic polypeptide precursor",

abstract = "The physiology of the incretin hormones, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), and their role in type 2 diabetes currently attract great interest. Recently we reported an essential role for prohormone convertase (PC) 1/3 in the cleavage of intestinal proglucagon, resulting in formation of GLP-1, as demonstrated in PC1/3-deficient mice. However, little is known about the endoproteolytic processing of the GIP precursor. This study investigates the processing of proGIP in PC1/3 and PC2 null mice and in cell lines using adenovirus-mediated overexpression. Supporting a role for PC1/3 in proGIP processing, we found co-localization of GIP and PC1/3 but not PC2 in intestinal sections by immunohistochemistry, and analysis of intestinal extracts from PC1/3-deficient animals demonstrated severely impaired processing to GIP, whereas processing to GIP was unaltered in PC2-deficient mice. Accordingly, overexpression of preproGIP in the neuroendocrine AtT-20 cell line that expresses high levels of endogenous PC1/3 and negligible levels of PC2 resulted in production of GIP. Similar results were obtained after co-expression of preproGIP and PC1/3 in GH4 cells that express no PC2 and only low levels of PC1/3. In addition, studies in GH4 cells and the alpha-TC1.9 cell line, expressing PC2 but not PC1/3, indicate that PC2 can mediate processing to GIP but also to other fragments not found in intestinal extracts. Taken together, our data indicate that PC1/3 is essential and sufficient for the production of the intestinal incretin hormone GIP, whereas PC2, although capable of cleaving proGIP, does not participate in intestinal proGIP processing and is not found in intestinal GIP-expressing cells.",

author = "Randi Ugleholdt and Poulsen, {Marie-Louise H} and Holst, {Peter J} and Jean-Claude Irminger and Cathrine Orskov and Jens Pedersen and Rosenkilde, {Mette M} and Xiaorong Zhu and Steiner, {Donald F} and Holst, {Jens Juul}",

note = "N{\o}gleord: Adenoviridae; Dyr, COS celler Cell Line; Cells, Kulturperler, Cercopithecus aethiops, Chromatography, Gel, Chromatography, High Pressure Liquid; cyklisk AMP, DNA, supplerer hinanden; dosis-respons sammenh{\ae}ng, Drug; Flowcytometri; Gastrisk Inhibitory Polypeptid; genetiske Vektorer, Glucose, Immunhistokemi, Insulin, Tarm, mus, mus, transgene; Modeller, genetiske, Peptider, polymerasek{\ae}dereaktion; Proprotein Convertase 1; Proprotein Convertases, RNA, RNA, Messenger, Radioimmunoassay, Rotter, receptorer, Mave-hormon; rekombinant Proteiner",

year = "2006",

doi = "10.1074/jbc.M601203200",

language = "English",

volume = "281",

pages = "11050--7",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

number = "16",

}

TY - JOUR

T1 - Prohormone convertase 1/3 is essential for processing of the glucose-dependent insulinotropic polypeptide precursor

AU - Ugleholdt, Randi

AU - Poulsen, Marie-Louise H

AU - Holst, Peter J

AU - Irminger, Jean-Claude

AU - Orskov, Cathrine

AU - Pedersen, Jens

AU - Rosenkilde, Mette M

AU - Zhu, Xiaorong

AU - Steiner, Donald F

AU - Holst, Jens Juul

N1 - Nøgleord: Adenoviridae; Dyr, COS celler Cell Line; Cells, Kulturperler, Cercopithecus aethiops, Chromatography, Gel, Chromatography, High Pressure Liquid; cyklisk AMP, DNA, supplerer hinanden; dosis-respons sammenhæng, Drug; Flowcytometri; Gastrisk Inhibitory Polypeptid; genetiske Vektorer, Glucose, Immunhistokemi, Insulin, Tarm, mus, mus, transgene; Modeller, genetiske, Peptider, polymerasekædereaktion; Proprotein Convertase 1; Proprotein Convertases, RNA, RNA, Messenger, Radioimmunoassay, Rotter, receptorer, Mave-hormon; rekombinant Proteiner

PY - 2006

Y1 - 2006

N2 - The physiology of the incretin hormones, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), and their role in type 2 diabetes currently attract great interest. Recently we reported an essential role for prohormone convertase (PC) 1/3 in the cleavage of intestinal proglucagon, resulting in formation of GLP-1, as demonstrated in PC1/3-deficient mice. However, little is known about the endoproteolytic processing of the GIP precursor. This study investigates the processing of proGIP in PC1/3 and PC2 null mice and in cell lines using adenovirus-mediated overexpression. Supporting a role for PC1/3 in proGIP processing, we found co-localization of GIP and PC1/3 but not PC2 in intestinal sections by immunohistochemistry, and analysis of intestinal extracts from PC1/3-deficient animals demonstrated severely impaired processing to GIP, whereas processing to GIP was unaltered in PC2-deficient mice. Accordingly, overexpression of preproGIP in the neuroendocrine AtT-20 cell line that expresses high levels of endogenous PC1/3 and negligible levels of PC2 resulted in production of GIP. Similar results were obtained after co-expression of preproGIP and PC1/3 in GH4 cells that express no PC2 and only low levels of PC1/3. In addition, studies in GH4 cells and the alpha-TC1.9 cell line, expressing PC2 but not PC1/3, indicate that PC2 can mediate processing to GIP but also to other fragments not found in intestinal extracts. Taken together, our data indicate that PC1/3 is essential and sufficient for the production of the intestinal incretin hormone GIP, whereas PC2, although capable of cleaving proGIP, does not participate in intestinal proGIP processing and is not found in intestinal GIP-expressing cells.

AB - The physiology of the incretin hormones, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), and their role in type 2 diabetes currently attract great interest. Recently we reported an essential role for prohormone convertase (PC) 1/3 in the cleavage of intestinal proglucagon, resulting in formation of GLP-1, as demonstrated in PC1/3-deficient mice. However, little is known about the endoproteolytic processing of the GIP precursor. This study investigates the processing of proGIP in PC1/3 and PC2 null mice and in cell lines using adenovirus-mediated overexpression. Supporting a role for PC1/3 in proGIP processing, we found co-localization of GIP and PC1/3 but not PC2 in intestinal sections by immunohistochemistry, and analysis of intestinal extracts from PC1/3-deficient animals demonstrated severely impaired processing to GIP, whereas processing to GIP was unaltered in PC2-deficient mice. Accordingly, overexpression of preproGIP in the neuroendocrine AtT-20 cell line that expresses high levels of endogenous PC1/3 and negligible levels of PC2 resulted in production of GIP. Similar results were obtained after co-expression of preproGIP and PC1/3 in GH4 cells that express no PC2 and only low levels of PC1/3. In addition, studies in GH4 cells and the alpha-TC1.9 cell line, expressing PC2 but not PC1/3, indicate that PC2 can mediate processing to GIP but also to other fragments not found in intestinal extracts. Taken together, our data indicate that PC1/3 is essential and sufficient for the production of the intestinal incretin hormone GIP, whereas PC2, although capable of cleaving proGIP, does not participate in intestinal proGIP processing and is not found in intestinal GIP-expressing cells.

U2 - 10.1074/jbc.M601203200

DO - 10.1074/jbc.M601203200

M3 - Journal article

C2 - 16476726

SN - 0021-9258

VL - 281

SP - 11050

EP - 11057

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 16

ER -

Prohormone convertase 1/3 is essential for processing of the glucose-dependent insulinotropic polypeptide precursor

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