Secretion of phospholipase B from Saccharomyces cerevisiae

Witt, Wolfgang; Mertsching, Angelika; König, Ekkehard

doi:10.1016/0005-2760(84)90111-5

Cited by 39 publications

(32 citation statements)

References 16 publications

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“…Solid ammonium sulfate was added to 80% saturation (W/V) at 0 C. The pH of the solution was adjusted with 1 N aqueous solution of ammonia to 7.5. The solution was left overnight at 4 C, and then the precipitate was removed by centrifugation at 20;000 Â g for 90 min at 4 C. The supernatant was put on a phenyl-Sepharose CL-4B column (2:5 Â 44 cm) equilibrated with 50 mM Tris-HCl buffer (pH 7.5) containing 80% saturated ammonium sulfate. After the unretained materials were eluted out with the same buffer, elution was performed with 50 mM Tris-HCl buffer (pH 7.5) containing no ammonium sulfate.…”

Section: Methodsmentioning

confidence: 99%

“…6). PLBs from microbes, 4,[22][23][24] plant cells, 25) and animal cells 26) have also been reported to convert lyso-PC to PC as a minor activity.…”

Section: Acyltransferase Activitymentioning

confidence: 99%

“…PLB catalyzes hydrolytic cleavage of both the sn-1 and the sn-2 acylester bonds of glycerophospholipids, and has been found in microorganisms, [1][2][3][4][5] plants, 6) and animal tissues. 7,8) The phopholipid deacylating enzymes characterized so far from yeasts at the molecular level belong to the category of the PLB group.…”

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confidence: 99%

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Purification and Characterization of Phospholipase B fromCandida utilis

Fujino

Akiyama

Akaboshi

et al. 2006

Bioscience, Biotechnology, and Biochemistry

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Section: Methodsmentioning

confidence: 99%

“…6). PLBs from microbes, 4,[22][23][24] plant cells, 25) and animal cells 26) have also been reported to convert lyso-PC to PC as a minor activity.…”

Section: Acyltransferase Activitymentioning

confidence: 99%

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Purification and Characterization of Phospholipase B fromCandida utilis

Fujino

Akiyama

Akaboshi

et al. 2006

Bioscience, Biotechnology, and Biochemistry

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“…The highly glycosylated enzyme of about 220 kDa (73 kDa for the protein part, predicted from the sequence) is enriched in the yeast plasma membrane but was also found in the periplasmic space and in the culture supernatant. The lysophospholipase activity of Plb1p greatly exceeds the activity catalyzing the first step of hydrolysis; thus, lyso-phospholipids do not accumulate as intermediate products of Plb1p activity (2)(3)(4). In addition, this enzyme has transacylase activity, catalyzing the synthesis of phosphatidylcholine (PtdCho) 1 from two molecules of lyso-phosphatidylcholine.…”

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confidence: 99%

Characterization and Function in Vivo of Two Novel Phospholipases B/Lysophospholipases fromSaccharomyces cerevisiae

Merkel

Fido

Mayr

et al. 1999

Journal of Biological Chemistry

102

118

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The yeast genome contains two genes, designated as PLB2 and PLB3, that are 67% and 62% identical, respectively, to PLB1, which codes for a phospholipase B/lysophospholipase in yeast (Lee, S. K., Patton, J. L., Fido, M., Hines, L. K., Kohlwein, S. D., Paltauf, F., Henry, S. A., and Levin, D. E. (1994) J. Biol. Chem. 269, 19725-19730). Deletion and overexpression studies and in vivo and in vitro activity measurements suggest that both genes indeed code for phospholipases B/lysophospholipases. In cell free extracts of a plb1 plb2 plb3 triple mutant, no phospholipase B activity was detectable. Upon overexpression of PLB2 in a plb1 plb3 mutant background, phospholipase B activity was detectable in the plasma membrane, periplasmic space extracts and the culture supernatant. Similar to Plb1p, Plb2p appears to accept all major phospholipid classes, with a preference for acidic phospholipids including phosphatidylinositol 3,4-bisphosphate and phosphatidic acid. Consistent with a function as an extracellular lysophospholipase, PLB2 overexpression conferred resistance to lyso-phosphatidylcholine. Deletion of Plb2p function had no effect on glycerophosphoinositol or glycerophosphocholine release in vivo, in contrast to a deletion of Plb3p function, which resulted in a 50% reduction of phosphatidylinositol breakdown and glycerophosphoinositol release from the cells. In vitro, Plb3p hydrolyzes only phosphatidylinositol and phosphatidylserine and, to a lesser extent, their lysoanalogs. Plb3p activity in a plb1 plb2 mutant background was observed in periplasmic space extracts. Both Plb3p and Plb2p display transacylase activity in vitro, in the presence or absence, respectively, of detergent.Phospholipases B catalyze the hydrolytic cleavage of both acylester bonds of glycerophospholipids. Products of phospholipase B activity are fatty acids and water-soluble glycerophosphodiesters. In yeast, soluble degradation products are released to some extent into the culture medium, and are thus an indicator of cellular phospholipase B activity. The only acylester-hydrolyzing enzyme from yeast characterized so far at the molecular level is a phospholipase B encoded by the PLB1 gene (1). The highly glycosylated enzyme of about 220 kDa (73 kDa for the protein part, predicted from the sequence) is enriched in the yeast plasma membrane but was also found in the periplasmic space and in the culture supernatant. The lysophospholipase activity of Plb1p greatly exceeds the activity catalyzing the first step of hydrolysis; thus, lyso-phospholipids do not accumulate as intermediate products of Plb1p activity (2-4). In addition, this enzyme has transacylase activity, catalyzing the synthesis of phosphatidylcholine (PtdCho) 1 from two molecules of lyso-phosphatidylcholine. The physiological function of yeast phospholipase B is still unclear; neither disruption of the PLB1 gene nor its overexpression result in detectable growth phenotypes. However, the amount of glycerophosphocholine (GroPCho) and glycerophosphoethanolamine released into the culture su...

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“…Several PLB have been identified in various microorganisms (1)(2)(3)(4)(5)(6)(7)(8) and in plants (9) as well as in the brush border membrane of mature enterocytes from three animal species including guinea pig (10 -13), rat (14 -17), and rabbit (17,18). It was later found that intestinal PLB actually displays a broader substrate specificity, including diacylglycerol, monoacylglycerol (12), and retinyl esters (19).…”

mentioning

confidence: 99%

Ectopic Epididymal Expression of Guinea Pig Intestinal Phospholipase B

Delagebeaudeuf

Gassama‐Diagne

Nauze

et al. 1998

Journal of Biological Chemistry

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Guinea pig intestinal phospholipase B is a calciumindependent phospholipase hydrolyzing sequentially the acyl ester bonds at sn-2 and sn-1 positions of glycerophospholipids, promoting the formation of sn-glycero-3-phosphocholine from phosphatidylcholine. This 140-kDa glycoprotein from the brush border membrane of differentiated enterocytes contributes to lipid digestion as an ectoenzyme. The cDNA coding for guinea pig phospholipase B was revealed to be the homologue of AdRab-B, an mRNA appearing in rabbit upon intestine development. The sequence predicts a polypeptide of 1463 amino acids displaying four homologous repeats, two of them containing the lipase consensus sequence GXSXG. A 5-kilobase transcript was particularly abundant in mature ileal and jejunal enterocytes but was also detected in epididymis, where phospholipase B displayed a higher molecular mass (170 kDa versus 140 kDa in intestine), with no obvious evidence for enzyme activity. Trypsin treatment of phospholipase B immunoprecipitated from epididymal membranes reduced its size to 140 kDa, coinciding with the appearance of a significant phospholipase A 2 activity. The same results were obtained in COS cells transfected with phospholipase B cDNA. Since sn-glycero-3-phosphocholine present at high concentrations in seminal plasma mainly stems from epididymis, this suggests a possible role of phospholipase B in male reproduction. This novel localization also unravels a mechanism of phospholipase B activation by limited proteolysis involving either trypsin in the intestinal lumen or a trypsin-like endopeptidase in the male reproductive tract.

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Secretion of phospholipase B from Saccharomyces cerevisiae

Cited by 39 publications

References 16 publications

Purification and Characterization of Phospholipase B fromCandida utilis

Purification and Characterization of Phospholipase B fromCandida utilis

Characterization and Function in Vivo of Two Novel Phospholipases B/Lysophospholipases fromSaccharomyces cerevisiae

Ectopic Epididymal Expression of Guinea Pig Intestinal Phospholipase B

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