Endocytosis of Secreted Carboxyl Ester Lipase in a Syndrome of Diabetes and Pancreatic Exocrine Dysfunction

Torsvik, Janniche; Johansson, Bente B.; Dalva, Monica; Marie, Michaël; Fjeld, Karianne; Johansson, Stefan; Bjørkøy, Geir; Saraste, Jaakko; Njølstad, Pål R.; Molven, Anders

doi:10.1074/jbc.m114.574244

Cited by 40 publications

(62 citation statements)

References 54 publications

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“…After studying native CEL isolated from pancreatic juice, we asked whether O-glycosylation of the recombinant protein expressed in human cell lines would reflect ABO/FUT2 status of the host cells. We started by analyzing HEK293 cells stably transfected with a CEL construct containing 16 VNTR repeats (11,12) as this line is a commonly used system for expressing mammalian glycoproteins. Genotyping of the cells, performed as described previously (32), predicted they derived from a blood group O and FUT2-positive individual.…”

Section: Characterization Of the O-glycome Of Recombinant Celmentioning

confidence: 99%

“…No ABO blood group antigens were detected on the secreted CEL protein even though the cells were FUT2-positive. To verify that the detected O-glycans originated solely from the mucinous domain of CEL, we analyzed HEK293 cells expressing CEL-TRUNC, a construct in which a stop codon had been introduced immediately before the VNTR region (11,12). Only two glycan peaks were detected (Fig.…”

Section: Characterization Of the O-glycome Of Recombinant Celmentioning

confidence: 99%

“…Human pancreatic cancer cell lines (BxPC-3, MIA PaCa-2, PANC-1) and HeLa cells (CCL-2) were obtained from American Type Culture Collection (ATCC). HEK293 cells (Clontech) stably transfected with pcDNA3.1/V5-His plasmids containing either WT CEL, CEL-TRUNC, or no insert were the same as described previously (11,12). PANC-1, HeLa, and HEK293 cells were cultured in DMEM (Sigma-Aldrich) with 500 g Geneticin (G-418; Invitrogen), 10% FBS (Invitrogen), and 100 units/ml penicillin/streptomycin.…”

Section: Cell Culturesmentioning

confidence: 99%

“…The variable length of the CEL VNTR makes this gene and its protein product highly polymorphic in human populations. In particular, rare mutational events affecting the VNTR region cause an inherited syndrome of diabetes and pancreatic exocrine dysfunction (MODY8) (10), most likely because of protein aggregation and endoplasmic reticulum stress resulting from altered repeat sequences (11)(12)(13). Moreover, a recombined allele between the VNTR regions of CEL and the neighboring pseudogene CELP is associated with a significantly increased risk for chronic pancreatitis (14,15).…”

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confidence: 99%

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The mucinous domain of pancreatic carboxyl-ester lipase (CEL) contains core 1/core 2 O-glycans that can be modified by ABO blood group determinants

Jellas

Johansson

Fjeld

et al. 2018

Journal of Biological Chemistry

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Carboxyl-ester lipase (CEL) is a pancreatic fat-digesting enzyme associated with human disease. Rare mutations in the CEL gene cause a syndrome of pancreatic exocrine and endocrine dysfunction denoted MODY8, whereas a recombined CEL allele increases the risk for chronic pancreatitis. Moreover, CEL has been linked to pancreatic ductal adenocarcinoma (PDAC) through a postulated oncofetal CEL variant termed feto-acinar pancreatic protein (FAPP). The monoclonal antibody mAb16D10 was previously reported to detect a glycotope in the highly O-glycosylated, mucin-like C terminus of CEL/FAPP. We here assessed the expression of human CEL in malignant pancreatic lesions and cell lines. CEL was not detectably expressed in neoplastic cells, implying that FAPP is unlikely to be a glycoisoform of CEL in pancreatic cancer. Testing of the mAb16D10 antibody in glycan microarrays then demonstrated that it recognized structures containing terminal GalNAc-␣1,3(Fuc-␣1,2)Gal (blood group A antigen) and also repeated protein sequences containing GalNAc residues linked to Ser/Thr (Tn antigen), findings that were supported by immunostainings of human pancreatic tissue. To examine whether the CEL glycoprotein might be modified by blood group antigens, we used high-sensitivity MALDI-TOF MS to characterize the released O-glycan pool of CEL immunoprecipitated from human pancreatic juice. We found that the O-glycome of CEL consisted mainly of core 1/core 2 structures with a composition depending on the subject's FUT2 and ABO gene polymorphisms. Thus, among digestive enzymes secreted by the pancreas, CEL is a glycoprotein with some unique characteristics, supporting the view that it could serve additional biological functions to its cholesteryl esterase activity in the duodenum.

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Section: Characterization Of the O-glycome Of Recombinant Celmentioning

confidence: 99%

Section: Characterization Of the O-glycome Of Recombinant Celmentioning

confidence: 99%

Section: Cell Culturesmentioning

confidence: 99%

mentioning

confidence: 99%

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The mucinous domain of pancreatic carboxyl-ester lipase (CEL) contains core 1/core 2 O-glycans that can be modified by ABO blood group determinants

Jellas

Johansson

Fjeld

et al. 2018

Journal of Biological Chemistry

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“…Ассоциацию с MODY8 и экзокринной дисфункцией ПЖ показали делеции в области VNTR (variable number of tandem repeats) гена CEL, вызывающие сдвиг рамки считывания в гене CEL (1686delT, 1785delC). Кроме того, очень короткий вариант аллеля VNTR, содержащий только 3 повтора (среднее количество повторов в популяции варьирует от 7 до 23), также оказался связанным с развитием MODY8, но предположено, что он имеет меньшую пене-трантность в отношении развития MODY [56]. Механизм развития этой формы СД неизвестен.…”

Section: терапевтический архив 4 2016unclassified

Molecular genetics of maturity-onset diabetes of the young

Воевода¹,

Иванова²,

Shakhtshneider³

et al. 2016

Ter. arkh.

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To verify the type of diabetes mellitus (DM) remains an extremely important problem in endocrinology, as along with types 1 and 2 DM there are rarer hereditary types of DM, including maturity-onset diabetes of the young (MODY). The latter is a genetic type of DM, which is characterized by an autosomal dominant inheritance. Eleven types of MODY (MODY 1 to MODY13) are identified; each is associated with mutations in the certain gene: HNF4A, GCK, HNF1A, PDX1, HNF1B, NEUROD1, KLF11, CEL, PAX4, INS, BLK, KCNJ11 and ABCC8. A molecular genetic testing for suspected MODY is conducted to verify the diagnosis and to define a subtype of MODY, patient management tactics, to predict the outcome of the disease and its complications in relation to the found subtype of MODY. It is also important to seek mutation causing MODY in terms of the early detection of MODY in the first-degree relatives of a proband, appropriate therapy of the disease, and prevention of its complications.

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Single nucleotide polymorphisms in CEL‐HYB1 increase risk for chronic pancreatitis through proteotoxic misfolding

et al. 2020

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Genetic variants contribute to the risk of chronic pancreatitis (CP) in adults and children. The risk variant CEL-HYB1, a recombinant hybrid allele of CEL and its neighboring pseudogene (CELP), encodes a pathogenic variant of the pancreatic digestive enzyme carboxyl ester lipase (CEL). We previously identified combinations of two non-synonymous SNPs, c.1463T>C (p. Ile488Thr) and c.1643C>T (p. Thr548Ile), in the break point region of CEL-HYB1. Herein, we tested whether these missense variants alter CP risk and their impact on functional properties of the CEL-HYB1 protein. Examination of CEL-HYB1 haplotypes in European patients and controls revealed that the combinationThr488-Ile548 was present only in cases (p ≤ .001). The lipase activity of purified recombinant CEL-HYB1 variants showed normal or near normal activity. CEL-HYB variants expressed in HEK293T cells all had decreased secretion compared with CEL, formed intracellular protein aggregates, and triggered endoplasmic reticulum stress. Thus, we propose that the presence of missense variants in CEL-HYB increases the pathogenicity of CEL-HYB1 through misfolding and gain-of-function proteotoxicity. Interestingly, Thr488-Ile548 and Thr488-Thr548 were equally pathogenic in the functional assays even though only the Thr488-Ile548 haplotype was significantly enriched in cases. The explanation for the mismatch between genetic and functional data requires further investigation.

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Endocytosis of Secreted Carboxyl Ester Lipase in a Syndrome of Diabetes and Pancreatic Exocrine Dysfunction

Cited by 40 publications

References 54 publications

The mucinous domain of pancreatic carboxyl-ester lipase (CEL) contains core 1/core 2 O-glycans that can be modified by ABO blood group determinants

The mucinous domain of pancreatic carboxyl-ester lipase (CEL) contains core 1/core 2 O-glycans that can be modified by ABO blood group determinants

Molecular genetics of maturity-onset diabetes of the young

Single nucleotide polymorphisms in CEL‐HYB1 increase risk for chronic pancreatitis through proteotoxic misfolding

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