Control of pancreatic bile-salt-dependent-lipase secretion by the glucose-regulated protein of 94kDa (Grp94)

Nganga, Alain; Bruneau, Nadine; Sbarra, Véronique; Lagadic‐Gossmann, Dominique; Petit-Thévenin, Josette Le

doi:10.1042/bj3520865

Cited by 19 publications

(10 citation statements)

References 54 publications

(76 reference statements)

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“…Such methodology is limited by the possibility that observed effects consecutive to amino acid substitution could be the result of an altered enzyme structure leading to a loss of function. This may not be the case as the mutagenized enzyme is normally secreted, whereas we have shown previously (20,21) that the unfolded or incorrectly folded BSDL is directed toward the ubiquitination and proteasome degradation pathway instead of the secretion route. Furthermore, the wild-type and mutagenized enzymes are still active to the same level on soluble substrates in the absence of activating bile salts.…”

Section: Fig 4 Secretion Of Y427s and Y453s Mutagenized Bsdl By Tramentioning

confidence: 68%

“…Those data suggest that substitutions significantly affect the enzyme activity on 4-NPC when recorded in the presence of activating concentrations of bile salt (4 mM NaTC). We have shown that unfolded BSDL cannot be secreted and instead is degraded by the proteasome (20,21). No degradation product can be detected on Western blotting performed on MC9 clone lysates (not shown).…”

Section: Resultsmentioning

confidence: 99%

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Site-directed Mutagenesis of the Distal Basic Cluster of Pancreatic Bile Salt-dependent Lipase

Aubert-Jousset¹,

Sbarra

Lagadic‐Gossmann

2004

Journal of Biological Chemistry

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Previous studies have postulated the presence of two bile salt-binding sites regulating the activity of the pancreatic bile salt-dependent lipase. One of these sites, located in an N-terminal basic cluster, has been identified as the specific bile salt-binding site. Interaction of primary bile salts with this proximal site induces the formation of a micellar binding site from a pre-existing nonspecific or pre-micellar bile salt-binding site. Here we have investigated the functional significance of another basic cluster comprised of amino acid residues Arg 423 , Lys 429 , Arg 454 , Arg 458 , and Lys 462 , distal from the catalytic site. For this purpose these residues were mutagenized in Ile or Ala residues. The mutagenized enzyme lost activity on both soluble and emulsified substrates in the presence of bile salts. However, in the absence of bile salts, the mutagenized enzyme displayed the same activity on soluble substrate as the wild-type recombinant enzyme. Consequently, the distal basic cluster may represent the nonspecific (or pre-micellar) bile salt-binding site susceptible to accommodate primary and secondary bile salts. According to the literature, tyrosine residue(s) should participate in this site. Therefore, two tyrosine residues, Tyr 427 and Tyr 453 , associated with the distal basic cluster were also mutagenized. Each tyrosine substitution to serine did not inhibit the enzyme activity on soluble substrate, independently of the presence of primary or secondary bile salts. However, the enzyme activity on cholesteryl oleate solubilized in primary bile salt micelles was decreased by mutations substantiating that these residues are part of the nonspecific bile salt-binding site.

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Section: Fig 4 Secretion Of Y427s and Y453s Mutagenized Bsdl By Tramentioning

confidence: 68%

Section: Resultsmentioning

confidence: 99%

Site-directed Mutagenesis of the Distal Basic Cluster of Pancreatic Bile Salt-dependent Lipase

Aubert-Jousset¹,

Sbarra

Lagadic‐Gossmann

2004

Journal of Biological Chemistry

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“…Opposite to mAb16D10, the mAbJ28 recognizes pancreatic tumor, but it has no effect on tumor growth. 8 Therefore, mAb16D10 could be of interest as therapeutic agents for passive immunization as described for breast cancers (48), provided that these antibodies specifically recognize human pancreatic tumoral tissues in vivo. Moreover, we have shown that the 16D10 antigen was expressed in metastatic pancreatic tumor and in tumor from patient after a preoperative chemotherapy.…”

Section: Discussionmentioning

confidence: 99%

“…Antibodies The patented IgM mAb16D10 directed against O-glycosylated repeated COOH-terminal mucin domain of FAPP (23), the patented IgG mAb8H8 against BSDL, 8 and the polyclonal antibodies (pAb) L64 against BSDL/FAPP (17) were homemade. mAbJ28 specific for the fucosylated J28 glycotope carried by repeated COOH-terminal sequences of FAPP was a generous gift from Dr. M-J.…”

Section: Humantissuesmentioning

confidence: 99%

“…During its transport from the endoplasmic reticulum up to the trans-Golgi network, BSDL is associated with intracellular membranes by means of a complex that involves glycosphingolipids of rafts (5) and the 94-kDa glucose-related protein (6,7). This Grp 94 controls the late folding steps and the sorting of the active enzyme toward secretion (8). Once in the trans-Golgi network and after completion of glycosylation (9,10), the complex is released from membranes on the phosphorylation of the threonine residue at position 340 (11) by a protein kinase CKII (12,13).…”

Section: Introductionmentioning

confidence: 99%

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Monoclonal antibody 16D10 to the COOH-terminal domain of the feto-acinar pancreatic protein targets pancreatic neoplastic tissues

Benkoël

Bernard²,

Payan-Defais³

et al. 2009

Molecular Cancer Therapeutics

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We have shown that the 16D10 antigen located on the mucin-like COOH-terminal domain of the feto-acinar pancreatic protein (FAPP) is expressed at the surface of human pancreatic tumor cell lines such as SOJ-6 cell line. Furthermore, an in vivo study indicates that targeting this cell-membrane glycopeptide by the use of the monoclonal antibody (mAb) 16D10 inhibits the growth of SOJ-6 xenografts in nude mice. To validate the potential use of the mAb16D10 in immune therapy, this study examined the expression of 16D10 antigens at the surface of human pancreatic adenocarcinomas versus control tissues. We examined the reactivity of mAb16D10 and mAb8H8 with pancreatic ductal adenocarcinomas (PDAC) compared with controls by using immunohistochemistry and confocal laser scanning microscopy. mAb8H8 does react with control or nontumoral human pancreatic tissues. mAb16D10 has a strong and specific reactivity with PDAC and does not react with other cancers of epithelia or normal tissues tested. Notable, mAb16D10 mostly recognizes membrane of tumoral cells. Furthermore, mAb8H8 and mAb16D10 recognized a protein of 110 to 120 kDa in homogenates of nontumoral and tumoral human pancreatic tissues, respectively. This size correlates with that of FAPP or with that of the normal counterpart of FAPP, the so-called bile salt-dependent lipase. The results suggest that mAb16D10 presents a unique specificity against PDAC; consequently, it could be effective in immune therapy of this cancer. Furthermore, mAb16D10 and mAb8H8 pair might be useful for diagnosis purpose in discriminating tumoral from nontumoral human pancreatic tissues. [Mol Cancer Ther 2009;8(2):282 -91]

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Characterization of a VNTR polymorphism in the coding region of the CEL gene

2002

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Control of pancreatic bile-salt-dependent-lipase secretion by the glucose-regulated protein of 94kDa (Grp94)

Cited by 19 publications

References 54 publications

Site-directed Mutagenesis of the Distal Basic Cluster of Pancreatic Bile Salt-dependent Lipase

Site-directed Mutagenesis of the Distal Basic Cluster of Pancreatic Bile Salt-dependent Lipase

Monoclonal antibody 16D10 to the COOH-terminal domain of the feto-acinar pancreatic protein targets pancreatic neoplastic tissues

Characterization of a VNTR polymorphism in the coding region of the CEL gene

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