[4] Lipases and hydrolase fold

Schrag, Joseph D.; Cygler, Mirosław

doi:10.1016/s0076-6879(97)84006-2

Cited by 256 publications

(207 citation statements)

References 77 publications

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“…The amino acid sequence of CGI-58 protein is highly conserved from humans to rodents throughout its entire length, and contains a region homologous to the ␣/␤ hydrolase fold found in a large family of proteins. It is classified as a member of the esterase/lipase/thioesterase subfamily, based on the sequence of its catalytic triad (36). Importantly, CGI-58 was identified as a causal gene of CDS, a disorder characterized by the accumulation of abnormally large amounts of lipid droplets in several organs (32).…”

Section: Discussionmentioning

confidence: 99%

“…1A). It belongs to a large protein family defined by a ␣/␤ hydrolase fold (36), and contains sequences highly conserved among members of the esterase/lipase/thioesterase subfamily (36). However, as compared with the conserved sequence motif of lipases (LLGHSLGG), the serine residue that is part of the catalytic triad is replaced by asparagine in the corresponding region of CGI-58 (LLGHNLGG; underlined in Fig.…”

Section: Identification Of Cgi-58 As a Binding Partner Of Perilipinmentioning

confidence: 99%

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CGI-58 Interacts with Perilipin and Is Localized to Lipid Droplets

Yamaguchi

Omatsu

Matsushita

et al. 2004

Journal of Biological Chemistry

236

110

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Lipid droplets (LDs) are a class of ubiquitous cellular organelles that are involved in lipid storage and metabolism. Although the mechanisms of the biogenesis of LDs are still unclear, a set of proteins called the PAT domain family have been characterized as factors associating with LDs. Perilipin, a member of this family, is expressed exclusively in the adipose tissue and regulates the breakdown of triacylglycerol in LDs via its phosphorylation. In this study, we used a yeast two-hybrid system to examine the potential function of perilipin. We found direct interaction between perilipin and CGI-58, a deficiency of which correlated with the pathogenesis of Chanarin-Dorfman syndrome (CDS). Endogenous CGI-58 was distributed predominantly on the surface of LDs in differentiated 3T3-L1 cells, and its expression increased during adipocyte differentiation. Overexpressed CGI-58 tagged with GFP gathered at the surface of LDs and colocalized with perilipin. This interaction seems physiologically important because CGI-58 mutants carrying an amino acid substitution identical to that found in CDS lost the ability to be recruited to LDs. These mutations significantly weakened the binding of CGI-58 with perilipin, indicating that the loss of this interaction is involved in the etiology of CDS. Furthermore, we identified CGI-58 as a binding partner of ADRP, another PAT domain protein expressed ubiquitously, by yeast two-hybrid assay. GFP-CGI-58 expressed in non-differentiated 3T3-L1 or CHO-K1 cells was colocalized with ADRP, and the CGI-58 mutants were not recruited to LDs carrying ADRP, indicating that CGI-58 may also cooperate with ADRP.

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Section: Discussionmentioning

confidence: 99%

Section: Identification Of Cgi-58 As a Binding Partner Of Perilipinmentioning

confidence: 99%

CGI-58 Interacts with Perilipin and Is Localized to Lipid Droplets

Yamaguchi

Omatsu

Matsushita

et al. 2004

Journal of Biological Chemistry

236

110

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“…Cutinases show no interfacial activation and are, therefore, placed as intermediates between lipases and esterases rather than being categorized as classical lipases. [5][6][7] This functional versatility has made them a good alternative to classical lipases for industrial use in detergent products. 8 To function in detergent mixtures, an enzyme needs to retain its catalytic activity under the conditions used for washing, that is, temperatures up to 60 C, and the presence of anionic and nonionic surfactants.…”

Section: Introductionmentioning

confidence: 99%

Thermodynamic and structural investigation of the specific SDS binding of humicola insolens cutinase

et al. 2014

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The interaction of lipolytic enzymes with anionic surfactants is of great interest with respect to industrially produced detergents. Here, we report the interaction of cutinase from the thermophilic fungus Humicola insolens with the anionic surfactant SDS, and show the enzyme specifically binds a single SDS molecule under nondenaturing concentrations. Protein interaction with SDS was investigated by NMR, ITC and molecular dynamics simulations. The NMR resonances of the protein were assigned, with large stretches of the protein molecule not showing any detectable resonances. SDS is shown to specifically interact with the loops surrounding the catalytic triad with medium affinity (K a % 10 5 M 21 ). The mode of binding is closely similar to that seen previously for binding of amphiphilic molecules and substrate analogues to cutinases, and hence SDS acts as a substrate mimic. In addition, the structure of the enzyme has been solved by X-ray crystallography in its apo form and after cocrystallization with diethyl p-nitrophenyl phosphate (DNPP) leading to a complex with monoethylphosphate (MEP) esterified to the catalytically active serine. TheAbbreviations: AA, all-atom; AoC, Aspergillus oryzae cutinase; AOT, sodium bis(2-ethylhexyl) sulfosuccinate; cmc, critical micelle concentration; DEP, diethylphosphate; DNPP, diethyl p-nitrophenyl phosphate; EDTA, ethylene diamine tetraacetate; FsC, Fusarium solani cutinase; GcC, Glomerella cingulata cutinase; HiC, Humicola insolens cutinase; HSQC, heteronuclear singlequantum coherence; IPTG, isopropyl b-D-1-thiogalactopyranoside; ITC, isothermal titration calorimetry; MD, molecular dynamics; MEP, monoethylphosphate; MWCO, molecular weight cut-off; PAGE, polyacrylamide gel electrophoresis; RMSD, root mean square deviation; RMSF, root mean square fluctuation; TES, N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid; SANS, small-angle neutron scattering; SDS, sodium dodecyl sulfate.Additional Supporting Information may be found in the online version of this article.An interactive view is available in the electronic version of the article. enzyme has the same fold as reported for other cutinases but, unexpectedly, esterification of the active site serine is accompanied by the ethylation of the active site histidine which flips out from its usual position in the triad.

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“…The active site for lipases and esterases typically consists of a catalytic triad formed by Ser-His-(Asp/ Glu), which is common for most serine hydrolases (17). A feature specific for many lipases and esterases, however, is the ␣/␤-hydrolase fold consisting of a series of parallel ␤-sheets and a number of helices that flank the sheets on both sides (18,19). Most lipases contain a lid controlling the access of substrates to the hydrophobic active site.…”

mentioning

confidence: 99%

The Lipolytic Proteome of Mouse Adipose Tissue

Birner‐Gruenberger

Susani-Etzerodt

Waldhuber

et al. 2005

Molecular & Cellular Proteomics

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In animals, plant seeds, and fungi, excessive amounts of lipids are stored in the form of intracellular triacylglycerol (TG) 1 and steryl ester deposits. Mammals store TG in adipose tissue as the primary source of energy during periods of starvation and increased energy demand. The balance of lipid storage and mobilization is tightly regulated to ensure whole body energy homeostasis. Stored fat is mobilized by activation of lipolytic enzymes, which degrade adipose TG and cholesterol esters and release non-esterified (free) fatty acids into the circulation. Variation in the concentration of circulating free fatty acids is an established risk factor for the development of insulin resistance in type 2 diabetes and related disorders (1-4). Cholesteryl esters represent the stored esterified form of cholesterol, which is an important constituent of membranes and a precursor of bile acid and steroid hormones. TG lipases and cholesteryl esterases are key enzymes in lipid mobilization, and respective lipolytic activities have been described in various tissues. However, only some of them have been identified on the molecular level. Many characterized lipases are secretory proteins (5-8), whereas the hormonesensitive lipase (HSL) (9), the monoglyceride lipase (MGL) (10), and the triacylglycerol hydrolase (TGH) (11) are intracellular enzymes. HSL was shown to hydrolyze TG and cholesteryl esters and was thought to catalyze the rate-limiting step in TG hydrolysis in adipose tissue. Yet in HSL knock-out mice, TG hydrolysis in adipose tissue as well as cholesteryl ester hydrolysis in macrophages were not abolished (9). MGL is ubiquitously expressed and rather specific for monoacylglycerol hydrolysis (10). TGH appears to be involved in TG mobilization in liver and is also expressed in adipose tissue where its role is not clear (11,12).The present study was aimed at the molecular identification of the lipolytic proteome of mouse adipose tissue using activity labels designed in our laboratory (Fig. 1). Basically an activity label is a molecule consisting of (i) a recognition site targeting a certain enzyme species, (ii) a properly positioned reactive site that forms a covalent bond with the target, and (iii) a tag for visualization and/or purification of the covalently bound target (13-16). The active site for lipases and esterases typically consists of a catalytic triad formed by Ser-His-(Asp/ Glu), which is common for most serine hydrolases (17). A feature specific for many lipases and esterases, however, is the ␣/␤-hydrolase fold consisting of a series of parallel ␤-sheets and a number of helices that flank the sheets on both sides (18,19). Most lipases contain a lid controlling the access of substrates to the hydrophobic active site. The same structural features are found in esterases except for the lid. Therefore, in contrast to lipases, most esterases do not show activation at lipid/water interfaces. However, it has to be emphasized that the borderline between lipases and esterFrom the ‡Institute

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[4] Lipases and hydrolase fold

Cited by 256 publications

References 77 publications

CGI-58 Interacts with Perilipin and Is Localized to Lipid Droplets

CGI-58 Interacts with Perilipin and Is Localized to Lipid Droplets

Thermodynamic and structural investigation of the specific SDS binding of humicola insolens cutinase

The Lipolytic Proteome of Mouse Adipose Tissue

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