Common domain structure of Ca<sup>2+</sup> and lipid‐binding proteins

Geisow, Michael J.

doi:10.1016/0014-5793(86)81445-4

Cited by 143 publications

(53 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We also examined the N-terminal region and found that both S47 and D166 catalytic dyad residues located within the ␣ -␤ hydrolase fold are critical for TAG hydrolysis within cells and in vitro. Within the proximal N-terminal region, we identifi ed two overlapping motifs, a glycine-rich motif and an amphipathic ␣ -helix that are predicted by sequence analysis to be neutral lipid substrate-binding domains ( 16,17 ). We show by site-directed mutational analysis that this region, especially the amphipathic ␣ -helix, is critical for TAG hydrolysis.…”

Section: Confocal Microscopymentioning

confidence: 93%

“…5F ), suggesting that the G14 residue may play an indirect role in TAG hydrolysis in living cells, possibly by participating in interactions with other cellular proteins. The second motif consists of the octapeptide FL G V YHIG, corresponding to amino acids [17][18][19][20][21][22][23][24], that is a nonconsensus version of the sequence FL X L XXXn, where X = any residue except proline and n = a nonpolar residue ( Fig. 5C ).…”

Section: Immunoblottingmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of desnutrin functional domains: critical residues for triacylglycerol hydrolysis in cultured cells

Duncan

Wang

Ahmadian

et al. 2010

Journal of Lipid Research

View full text Add to dashboard Cite

Triacylglycerol (TAG) serves as the major energy reserve in higher eukaryotic organisms ( 1 ). FFAs are mobilized from TAG through the hydrolytic action of lipases to provide substrates for oxidative metabolism in tissues, as well as substrates for synthesis of complex lipids and signaling molecules. Lipolysis, the breakdown of TAG to FFA and glycerol, occurs in an orderly and regulated manner, with different enzymes acting sequentially at each step ( 1-3 ). Hydrolysis of TAG to diacylglycerol is the fi rst step ( 4-6 ), and our laboratory, and others, recently identifi ed a novel TAG hydrolase in mice that we called desnutrin ( 5 ) [also known as TTS2.2, patatin-like phospholipase A domain containing 2, iPLA ( 4 ), or in humans, adipose triglyceride lipase (ATGL) ( 6 )]. Desnutrin is highly expressed in white and brown adipose tissue, where it is a major TAG lipase, but is also found at lower levels in most other tissues where it also plays an important role in TAG hydrolysis ( 5,7,8 ).Desnutrin contains an N-terminal patatin-like domain, spanning amino acids 8-180, that is characteristic of many plant lipid acyl hydrolases ( 5, 9 ). Enzymatic activity of desnutrin is predicted to be derived from an S47-D166 catalytic dyad that lies within an ␣ -␤ hydrolase fold in the patatin-like domain ( 5, 9 ). Mutation of the S47 residue to alanine has been shown to result in a complete loss of function in vitro, confi rming the predicted serine-esterase activity of this enzyme ( 10 ). However, the requirement of the D166 site has not yet been tested. Four individuals with point mutations in desnutrin/ATGL have been identifi ed, and all developed neutral lipid storage disease with myopathy (NLSDM) ( 11,12 ). In one, a duplication mutation within the N-terminal patatin-like domain caused a frameshift after L159 that is predicted to truncate the enzyme at amino acid position 178 as well as to cause a D166R mutaAbstract Murine desnutrin/human ATGL is a triacylglycerol (TAG) hydrolase with a predicted catalytic dyad within an ␣ -␤ hydrolase fold in the N-terminal region. In humans, mutations resulting in C-terminal truncation cause neutral lipid storage disease with myopathy. To identify critical functional domains, we measured TAG breakdown in cultured cells by mutated or truncated desnutrin. In vitro, C-terminally truncated desnutrin displayed an even higher apparent V max than the full-length form without changes in K m , which may be explained by our fi nding of an interaction between the C-and N-terminal domains. In live cells, however, C-terminally truncated adenoviral desnutrin had lower TAG hydrolase activity. We investigated a role for the phosphorylation of C-terminal S406 and S430 residues but found that these were not necessary for TAG breakdown or lipid droplet localization in cells. The predicted N-terminal active sites, S47 and D166, were both critical for TAG hydrolysis in live cells and in vitro. We also identifi ed two overlapping N-terminal motifs that predict lipid substrate binding domains, a glycine-rich ...

show abstract

Section: Confocal Microscopymentioning

confidence: 93%

Section: Immunoblottingmentioning

confidence: 99%

Characterization of desnutrin functional domains: critical residues for triacylglycerol hydrolysis in cultured cells

Duncan

Wang

Ahmadian

et al. 2010

Journal of Lipid Research

View full text Add to dashboard Cite

show abstract

“…Some key residues in this site have been identified by site-directed mutagenesis of human factor IX (Handford et al, 1991). A further calcium-binding unit, the endonexin fold, has been identified in such proteins as calelectrin and lipocortin (Geisow, 1986), and involves up to 70 amino acid residues in a calcium-dependent phosphate-binding site. Although the calcium-binding sites of these groups are not fully characterized they are unlike the repeats of the ARP.…”

Section: Transcription and Translation Of Artp In Vitvo And Import Inmentioning

confidence: 99%

A novel calcium‐binding protein from Euglena gracilis

Gumpel

Smith

1992

European Journal of Biochemistry

View full text Add to dashboard Cite

We have isolated a novel cDNA from Euglena gracilis that encodes a protein composed of 24.9% aspartate with an estimated pZ of 3.56, and a deduced molecular mass of 73 542 Da. The first 20 or so amino acids are hydrophobic and resemble a signal sequence. The rest of the polypeptide is composed of a 23-amino-acid repeat. There are 30 repeats, of which 23 are full length. Part of the consensus sequence derived from the repeats has some similarity to the loop of the EF-hand type calcium-binding motif. Evidence is presented that a fusion protein of t h s novel protein with /I-galactosidase can bind calcium. Northern blotting indicates a single transcript of 2.3 kb (the same size as the cDNA). In-vitro translation of the cDNA gives a protein that migrates on SDSjPAGE with an apparent molecular mass of 120 -125 kDa. The protein is processed into a smaller, proteaseprotected form (1 10-120 kDa) when translated in the presence of canine pancreatic microsomal vesicles. This suggests that the protein is targeted across the endoplasmic reticulum membrane in vivo, and is the first report of a signal sequence from E. gracilis. We propose that the cDNA obtained encodes a novel calcium-binding protein that is either secreted or resident in the endomembrane system of E. gracilis, and call it the acidic-repeat protein.

show abstract

“…Calelectrin was initially described in the electric organ of Torpedo marmorata and later in mammalian tissues, as a protein able to bind calcium and chromaffin granule membranes, and was considered as a candidate for Ca 2+ dependent Correspondence address: H. van den Bosch, Biochemistry Laboratory, Padualaan 8, 3584 CH Utrecht, The Netherlands regulators of membrane events [11][12][13]. The lipocortins, the calpactins and the calelectrins all possess a 17 amino acid residue consensus sequence [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Lipocortin inhibition of extracellular and intracellular phospholipases A₂ is substrate concentration dependent

Aarsman¹,

Mynbeek²,

Bosch³

et al. 1987

FEBS Letters

113

View full text Add to dashboard Cite

Hydrolysis of Escherichia coli membrane phospholipids by pancreatic phospholipase A 2 was inhibited by lipocortin from human monocytes in a substrate dependent manner. Inhibition was completely overcome at substrate concentrations above 250 pM. Lipocortin also inhibited partially purified preparations of two intracellular phospholipases A 2 isolated from rat liver mitochondria and rat platelets when these enzymes were assayed at low micromolar concentrations of phosphatidylethanolamine. Inhibition gradually decreased with increasing substrate concentrations both for pancreatic and platelet phospholipase A 2 and became completely abolished above 15 and 50/tM phosphatidylethanolamine, respectively.

show abstract

Common domain structure of Ca²⁺ and lipid‐binding proteins

Cited by 143 publications

References 32 publications

Characterization of desnutrin functional domains: critical residues for triacylglycerol hydrolysis in cultured cells

Characterization of desnutrin functional domains: critical residues for triacylglycerol hydrolysis in cultured cells

A novel calcium‐binding protein from Euglena gracilis

Lipocortin inhibition of extracellular and intracellular phospholipases A₂ is substrate concentration dependent

Contact Info

Product

Resources

About

Common domain structure of Ca2+ and lipid‐binding proteins

Cited by 143 publications

References 32 publications

Characterization of desnutrin functional domains: critical residues for triacylglycerol hydrolysis in cultured cells

Characterization of desnutrin functional domains: critical residues for triacylglycerol hydrolysis in cultured cells

A novel calcium‐binding protein from Euglena gracilis

Lipocortin inhibition of extracellular and intracellular phospholipases A2 is substrate concentration dependent

Contact Info

Product

Resources

About

Common domain structure of Ca²⁺ and lipid‐binding proteins

Lipocortin inhibition of extracellular and intracellular phospholipases A₂ is substrate concentration dependent