A structural domain (the lid) found in pancreatic lipases is absent in the guinea pig (phospho)lipase

Hjorth, A.; Carrière, Frédéric; Cudrey, Claire; Wöldike, H.; Boel, Esper; Lawson, David M.; Ferrato, Francine; Cambillau, Christian; Dodson, Guy

doi:10.1021/bi00069a003

Cited by 179 publications

(142 citation statements)

References 33 publications

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“…In this report we describe the characterization of one of them, nmd, encoding an unknown member of the enzyme family of lipases. The predicted NMD polypeptide lacks a so-called 'lid' domain, as was earlier described for guinea pig pancreatic (phospho)lipase GPLRP2 and the phospholipase Al from hornet venom (Dolml) [5,6].…”

Section: Introductionmentioning

confidence: 85%

“…Sequence analysis revealed that nmd is a novel gene encoding a 50 kDa protein, which probably represents a new member of the enzyme family of lipases [5,18,19]. The similarity with other lipases is sufficiently high to classify NMD as an a/ (3-hydrolase-fold enzyme [20,21].…”

Section: Discussionmentioning

confidence: 99%

“…In the absence of an aggregated lipid substrate, the lid domain prevents access to the active site. In the presence of water-insoluble substances, the lid domain as well as another surface loop (the so-called (35 loop) undergo large conformational changes thus opening access to the active site and creating the oxyanion hole [5,19,21]. In GPLRP2, containing a 'mini-lid', the catalytic site is freely accessible, and it displays a high phospholipase activity [5,6].…”

Section: Discussionmentioning

confidence: 99%

“…In the presence of water-insoluble substances, the lid domain as well as another surface loop (the so-called (35 loop) undergo large conformational changes thus opening access to the active site and creating the oxyanion hole [5,19,21]. In GPLRP2, containing a 'mini-lid', the catalytic site is freely accessible, and it displays a high phospholipase activity [5,6]. In the pancreatic (phospho)lipase of the coypu (coypu pancreatic lipaserelated protein 2 or CoPLRP2), the lid domain is not deleted, but the stabilizing interactions observed in classical pancreatic lipase between the lid domain, the protein core and the (35 loop, are missing.…”

Section: Discussionmentioning

confidence: 99%

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nmd, a novel gene differentially expressed in human melanoma cell lines, encodes a new atypical member of the enzyme family of lipases

et al. 1997

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nmd, a novel gene, was isolated by applying the differential mRNA display method to human melanoma cell lines with different metastatic capacity. In a panel of 17 other human tumor cell lines, nmd RNA expression could only be detected at low levels in T24 (bladder carcinoma) and Caco-2 (colon adenocarcinoma). Furthermore, it was found in placenta and liver, but not in skin, colon, spleen, lung, muscle, prostate and kidney. Sequence analysis classified the nmd gene product as a new member of the enzyme family of lipases (almost 30% identity in amino acid sequence with other human lipases). Active site residues of lipases were conserved in NMD, but NMD lacks the regulatory lid domain, which controls entry to the active site in classical lipases. A similar deletion was earlier reported by others in the guinea pig pancreatic (phospho)lipase GPLRP2 and the phospholipase Al from hornet venom (Dolml).

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Section: Introductionmentioning

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

nmd, a novel gene differentially expressed in human melanoma cell lines, encodes a new atypical member of the enzyme family of lipases

et al. 1997

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“…This is a consequence of the phenomenon called interfacial activation 17 . In most cases, the active site of the lipase is covered by short amphiphathic α-helix called the lid (closed form) make it inaccessible in the medium 18 .…”

Section: Introductionmentioning

confidence: 99%

Methacrylated Chitosan Based UV Curable Support for Enzyme Immobilization

Bayramoğlu

2017

Mat. Res.

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UV curing is simple, fast and effective procedure for enzyme immobilization with minimized enzymatic activity. UV-curable methacrylated chitosan is here proposed as a support material for immobilization of lipase. The morphology of the polymeric support was characterized by scanning electron microscopy (SEM). Both covalently (CIL) and physically (PIL) immobilized enzyme is analyzed in terms of enzymatic activity as a function of reusability, pH, storage, as well as stability under various experimental conditions. The recovery of activity of lipases immobilized onto a photocrosslinked polymer network was 82.0% and 70.0% for CIL and PIL, respectively. The optimum pH value for the free lipase was at pH 6.0. The optimum pH of the both CIL and PIL was shifted to pH 7.0. Immobilization increased the thermostability of the lipase from 50 ºC to 62 ºC. The free enzyme lost all its activity within 15 days. Repeated batch experiments show that about 61% of the enzyme activity of CIL and 41% of PIL was retained after 8 cycles.

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Reactivation of the totally inactive pancreatic lipase RP1 by structure-predicted point mutations

Roussel

Bezzine

et al. 1998

Proteins

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Both classical pancreatic lipase (DPL) and pancreatic lipase-related protein 1 (DPLRP1) have been found to be secreted by dog exocrine pancreas. These two proteins were purified to homogeneity from canine pancreatic juice and no significant catalytic activity was observed with dog PLRP1 on any of the substrates tested: di- and tri-glycerides, phospholipids, etc. DPLRP1 was crystallized and its structure solved by molecular replacement and refined at a resolution of 2.10 A. Its structure is similar to that of the classical PL structures in the absence of any inhibitors or micelles. The lid domain that controls the access to the active site was found to have a closed conformation. An amino-acid substitution (Ala 178 Val) in the DPLRP1 may result in a steric clash with one of the acyl chains observed in the structures of a C11 alkyl phosphonate inhibitor, a transition state analogue, bound to the classical PL. This substitution was suspected of being responsible for the absence of DPLRP1 activity. The presence of Val and Ala residues in positions 178 and 180, respectively, are characteristic of all the known PLRP1, whereas Ala and Pro residues are always present in the same positions in all the other members of the PL gene family. Introducing the double mutation Val 178 Ala and Ala 180 Pro into the human pancreatic RP1 (HPLRP1) gene yielded a well expressed and folded enzyme in insect cells. This enzyme is kinetically active on triglycerides. Our findings on DPLRP1 and HPLRP1 are therefore likely to apply to all the RP1 lipases.

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A structural domain (the lid) found in pancreatic lipases is absent in the guinea pig (phospho)lipase

Cited by 179 publications

References 33 publications

nmd, a novel gene differentially expressed in human melanoma cell lines, encodes a new atypical member of the enzyme family of lipases

nmd, a novel gene differentially expressed in human melanoma cell lines, encodes a new atypical member of the enzyme family of lipases

Methacrylated Chitosan Based UV Curable Support for Enzyme Immobilization

Reactivation of the totally inactive pancreatic lipase RP1 by structure-predicted point mutations

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