Experimental Mycology最新文献

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Electrophoretic Detection of Cytoplasmic Serine Proteinases (Gelatinases) in Candida albicans 白色念珠菌细胞质丝氨酸蛋白酶(明胶酶)的电泳检测
Experimental Mycology Pub Date : 1994-09-01 DOI: 10.1006/EMYC.1994.1025
M. Rodier, B. Moudni, M. Ghazali, C. Lacroix, J. Jacquemin
{"title":"Electrophoretic Detection of Cytoplasmic Serine Proteinases (Gelatinases) in Candida albicans","authors":"M. Rodier, B. Moudni, M. Ghazali, C. Lacroix, J. Jacquemin","doi":"10.1006/EMYC.1994.1025","DOIUrl":"https://doi.org/10.1006/EMYC.1994.1025","url":null,"abstract":"Abstract Rodier, M.-H., El Moudni, B., Ghazali, M., Lacroix, C., and Jacquemin, J.-L. 1994. Electrophoretic detection of cytoplasmic serine proteinases (gelatinases) in Candida albicans. Experimental Mycology 18: 267-270. Proteolytic activities of Candida albicans were identified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis with gels containing gelatin as proteinase substrate. Cytoplasmic extracts of five isolates were tested with this method. Two major proteolytic activities of Mr 50,000 and 60,000 were conserved between isolates whereas two other less intense activities of Mr 90,000 and 120,000 were only detected in three isolates. The two bands of proteolysis of Mr, 50,000 and 60,000 were revealed between pH 5.0 and pH 8.0, optimally active at pH 7.0, and their isoelectric point was 4.5. They were not influenced by the presence of 2-mercaptoethanol. Their sensitivity to phenylmethylsulfonyl fluoride and chymostatin allowed them to be characterized as serine proteinases. These two neutral proteinases were not secreted in culture supernatant.","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"1 1","pages":"267-270"},"PeriodicalIF":0.0,"publicationDate":"1994-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75553380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Spore Dormancy Mutants of Phycomyces 植菌属孢子休眠突变体
Experimental Mycology Pub Date : 1994-09-01 DOI: 10.1006/emyc.1994.1022
Francisco Rivero, Enrique Cerdá-Olmedo
{"title":"Spore Dormancy Mutants of Phycomyces","authors":"Francisco Rivero,&nbsp;Enrique Cerdá-Olmedo","doi":"10.1006/emyc.1994.1022","DOIUrl":"10.1006/emyc.1994.1022","url":null,"abstract":"<div><p>Rivero, F., and Cerdá-Olmedo, E. 1994. Spore dormancy mutants of <em>Phycomyces. Experimental Mycology</em> 18, 221-229. The spores of the Zygomycete <em>Phycomyces blakesleeanus</em> are called dormant because few of them germinate when placed in a medium that sustains mycelial growth and development. Nearly all the spores germinate after activation, that is, exposure to heat or certain chemicals. We have looked for mutants whose spores would not need activation. Nine mutants formed authentic, but transient spores, which germinated spontaneously in the sporangium. Mutant mycelia had lower alcohol and aldehyde dehydrogenase activities and less glycogen than wild-type mycelia. The spontaneous germination and the metabolic alterations are attributed to the same recessive mutations. No differences were found between mutants and wild type in the cyclic AMP and fructose 2,6-bisphosphate concentrations in immature sporangia and the trehalase activity in the mycelia. In another mutant the spore primordia did not form spores, but remained viable for some time in the sporangium. The mutants were difficult to keep in the laboratory (except as lyophils); this stresses the importance of preventing spore germination in the sporangium.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 3","pages":"Pages 221-229"},"PeriodicalIF":0.0,"publicationDate":"1994-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1022","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75744146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
Electrophoretic Detection of Cytoplasmic Serine Proteinases (Gelatinases) in Candida albicans 白色念珠菌细胞质丝氨酸蛋白酶(明胶酶)的电泳检测
Experimental Mycology Pub Date : 1994-09-01 DOI: 10.1006/emyc.1994.1025
Marie-Helene Rodier, Brahim El Moudni, Machhour Ghazali, Catherine Lacroix, Jean-Louis Jacquemin
{"title":"Electrophoretic Detection of Cytoplasmic Serine Proteinases (Gelatinases) in Candida albicans","authors":"Marie-Helene Rodier,&nbsp;Brahim El Moudni,&nbsp;Machhour Ghazali,&nbsp;Catherine Lacroix,&nbsp;Jean-Louis Jacquemin","doi":"10.1006/emyc.1994.1025","DOIUrl":"https://doi.org/10.1006/emyc.1994.1025","url":null,"abstract":"<div><p>Rodier, M.-H., El Moudni, B., Ghazali, M., Lacroix, C., and Jacquemin, J.-L. 1994. Electrophoretic detection of cytoplasmic serine proteinases (gelatinases) in <em>Candida albicans. Experimental Mycology</em> 18: 267-270. Proteolytic activities of <em>Candida albicans</em> were identified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis with gels containing gelatin as proteinase substrate. Cytoplasmic extracts of five isolates were tested with this method. Two major proteolytic activities of <em>M</em><sub>r</sub> 50,000 and 60,000 were conserved between isolates whereas two other less intense activities of <em>M</em><sub>r</sub> 90,000 and 120,000 were only detected in three isolates. The two bands of proteolysis of <em>M</em><sub>r</sub>, 50,000 and 60,000 were revealed between pH 5.0 and pH 8.0, optimally active at pH 7.0, and their isoelectric point was 4.5. They were not influenced by the presence of 2-mercaptoethanol. Their sensitivity to phenylmethylsulfonyl fluoride and chymostatin allowed them to be characterized as serine proteinases. These two neutral proteinases were not secreted in culture supernatant.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 3","pages":"Pages 267-270"},"PeriodicalIF":0.0,"publicationDate":"1994-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1025","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90007756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
The Heterothallic Life Cycle of Agaricus bisporus var. burnettii and the Inheritance of Its Tetrasporic Trait 双孢蘑菇的异孢生活史及其四孢性状的遗传
Experimental Mycology Pub Date : 1994-09-01 DOI: 10.1006/emyc.1994.1020
Richard W. Kerrigan, Micheline Imbernon, Philippe Callac, Christophe Billette, Jean-Marc Olivier
{"title":"The Heterothallic Life Cycle of Agaricus bisporus var. burnettii and the Inheritance of Its Tetrasporic Trait","authors":"Richard W. Kerrigan,&nbsp;Micheline Imbernon,&nbsp;Philippe Callac,&nbsp;Christophe Billette,&nbsp;Jean-Marc Olivier","doi":"10.1006/emyc.1994.1020","DOIUrl":"10.1006/emyc.1994.1020","url":null,"abstract":"<div><p>Kerrigan, R. W., Imbernon, M., Callac, P., Billette, C., And Olivier, J.-M. 1994. The heterothallic life cycle of <em>Agaricus bisporus</em> var. <em>burnettii</em> and the inheritance of its tetrasporic trait. <em>Experimental Mycology</em> 18, 193-210. The new taxon <em>Agaricus bisporus</em> var. <em>burnettii</em> was recently proposed for a tetrasporic population of the species discovered in California. This tetrasporic variety is interfertile with the familiar bisporic variety of the species. In the present study we determined the heritability of parental basidial morphologies in 91 first-generation intervarietal hybrids descended from either of two genetically distinct strains of var. <em>burnettii</em>. Large samples of basidiospore offspring of one of these parental var. <em>burnettii</em> strains, and of one of the first-generation intervarietal hybrids, were evaluated with respect to their ploidy (n vs n + n). The life cycle of var. <em>burnettii</em> was deduced and the mating type alleles of the two var. <em>burnettii</em> parents stocks were determined. We found that: (1) The life cycle of this variety, of which more than 90% of the basidia were four-spored, was predominantly heterothallic. About 90% of the basidiospores were homokaryotic. (2) Its incompatibility system was unifactorial. Four different, previously unknown mating type alleles were found. (3) The tetrasporic trait was always transmitted to first-generation hybrids between bisporic and tetrasporic varieties. Of 91 intervarietal hybrids, 88 had a majority of four-spored basidia while 3 had predominantly three-spored basidia, and none had more than a small percentage of bisporic basidia. (4) The basidiospore offspring of these hybrids germinated normally, producing a high percentage of homokaryons. For one hybrid sporocarp this percentage was estimated at 77% by three methods (growth rate, mating competence, and multilocus allozyme genotype) which generally agreed with one another. These data shed new light on the genetics of <em>A. bisporus</em> and will be useful in related breeding work.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 3","pages":"Pages 193-210"},"PeriodicalIF":0.0,"publicationDate":"1994-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85356952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 72
Distribution of Chitin Synthetase and Various Membrane Marker Enzymes in Chitosomes and Other Organelles of the Slime Mutant of Neurospora crassa 粗神经孢子菌黏液突变体壳质体及其他细胞器中几丁质合成酶及各种膜标记酶的分布
Experimental Mycology Pub Date : 1994-06-01 DOI: 10.1006/emyc.1994.1018
Carlos A. Leal-Morales, Charles E. Bracker, Salomon Bartnicki-Garcia
{"title":"Distribution of Chitin Synthetase and Various Membrane Marker Enzymes in Chitosomes and Other Organelles of the Slime Mutant of Neurospora crassa","authors":"Carlos A. Leal-Morales,&nbsp;Charles E. Bracker,&nbsp;Salomon Bartnicki-Garcia","doi":"10.1006/emyc.1994.1018","DOIUrl":"10.1006/emyc.1994.1018","url":null,"abstract":"<div><p>Leal-Morales, C. A., Bracker, C. E., and Bartnicki-Garcia, S. 1994. Distribution of chitin synthetase and various membrane marker enzymes in chitosomes and other organelles of the slime mutant of <em>Neurospora crassa. Experimental Mycology</em> 18, 168-179. The subcellular distribution of chitin synthetase was assessed in cell homogenates of the slime mutant of <em>Neurospora crassa</em> . The activities of several marker enzymes were monitored to establish the relationship between organelles containing chitin synthetase and other cytoplasmic organelles. Before cell breakage, the plasma membrane of the slime mutant was radiolabeled with traces of [<sup>3</sup>H]concanavalin A. Two major (I, II) and two minor (III, IV) bands of particles containing chitin synthetase were detected after sedimentation of crude cell homogenates on a discontinuous sucrose gradient [1-4 h at 81,500g (<em>R</em><sub>av</sub>)]. The main population (I) consisted of microvesicles (chitosomes) which sedimented more slowly than most other membranous organelles. The other major population (II) of chitin synthetase particles cosedimented with the plasma membrane markers, [<sup>3</sup>H]concanavalin A and a vanadate-sensitive ATPase. Despite differences in sedimentation velocity, the two major chitin synthetase populations (I and II) eventually equilibrated at the same buoyant density (1.12 g/ml). One of the two minor bands (III) of chitin synthetase was associated with plasma membrane surface marker. Band IV, the smallest fraction with chitin synthetase activity was not characterized. A soluble, dialyzable, partially thermostable factor found in the cytosol of <em>N. crassa</em> enhanced the chitin synthetase activity of chitosomes (4.6 to 4.8-fold); to a lesser extent (about 2-fold) it also enhanced the activity of the other chitin synthetase populations. This stimulatory factor found in the upper portion of the gradients affected the levels of chitin synthetase activity detected, particularly that of chirpspines in peak I. By a combination of centrifugation procedures, the subcellular populations of chitin synthetase were clearly separated from other organelles with similar [endoplasmic reticulum, <em>d</em> = 1.118, marker: phosphatidyl choline glyceride transferase] or different equilibrium densities [vacuole, <em>d</em> = 1.170 g/ml, marker: α-mannosidase; mitochondria, <em>d</em> = 1.175, marker: succinate dehydrogenase]. The absence of markers for plasma membrane, mitochondria, and vacuoles from the chitosome population and the partial but unequivocal separation of the endoplasmic reticulum marker from chitosomal chitin synthetase provide additional verification that these microvesicles are distinct subcellular structures and not fragments of other organelles. The findings support the existence of a unique secretory pathway based on chitosome microvesicles as the main conveyors of chitin synthetase to the cell surface.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 2","pages":"Pages 168-179"},"PeriodicalIF":0.0,"publicationDate":"1994-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80132708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 12
Elongation Growth and Gravitropic Curvature in the Flammulina velutipes (Agaricales) Fruiting Body 金针菇子实体的伸长生长和向地弯曲
Experimental Mycology Pub Date : 1994-06-01 DOI: 10.1006/emyc.1994.1016
Eva Haindl, Jan Monzer
{"title":"Elongation Growth and Gravitropic Curvature in the Flammulina velutipes (Agaricales) Fruiting Body","authors":"Eva Haindl,&nbsp;Jan Monzer","doi":"10.1006/emyc.1994.1016","DOIUrl":"10.1006/emyc.1994.1016","url":null,"abstract":"<div><p>Haindt, E., and Monzer, J. 1994. Elongation growth and gravitropic curvature in the <em>Flammulina velutipes</em> (Agaricales) fruiting body. <em>Experimental Mycology</em> 18, 150-158. Differential elongation of stipe hyphae drives the gravitropic reorientation of <em>Flammulina velutipes</em> (Agaricales) fruiting bodies. The gravitropic curvature is strictly dependent on the presence of the transition zone between pileus and stipe. Elongation growth, providing the driving force for curvature, is also promoted by the pileus. Gravitropic curvature is successfully suppressed by clinostatic rotation, but the elongation rate is not affected. Explantation of fruiting body stipes lowers curvature and elongation rates corresponding to explant size reduction. In <em>Flammulina</em> , 25 mm length of transition zone explants is an efficient size for reproducible curvature and elongation during 48- to 72-h curvature tests. Submersion of specimens in aqueous medium causes cessation of the gravitropic curvature, but does not affect elongation. Thus the involvement of a diffusible factor in transmission of the curvature signal is probable. Splitting the fruiting body stipe in segments of <span><math><mtext>1</mtext><mtext>8</mtext></math></span> diameter does not suppress the gravitropic response, and the segments are individually reoriented to the vertical. It is concluded that the graviresponse of the <em>Flammulina</em> fruiting body is based on cellular perception of the gravistimulus and that a differential growth signal is transmitted in the stipe by a soluble factor that regulates hyphal elongation.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 2","pages":"Pages 150-158"},"PeriodicalIF":0.0,"publicationDate":"1994-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82351619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 14
The Electrostatic Nature of the Cell Surface of Candida albicans: A Role in Adhesion 白色念珠菌细胞表面的静电性质:在粘附中的作用
Experimental Mycology Pub Date : 1994-06-01 DOI: 10.1006/emyc.1994.1013
Lorraine Jones, Paul O'Shea
{"title":"The Electrostatic Nature of the Cell Surface of Candida albicans: A Role in Adhesion","authors":"Lorraine Jones,&nbsp;Paul O'Shea","doi":"10.1006/emyc.1994.1013","DOIUrl":"10.1006/emyc.1994.1013","url":null,"abstract":"<div><p>Jones, L., and O'Shea, P. 1994. The electrostatic nature of the cell surface of <em>Candida albicans:</em> A role in adhesion. <em>Experimental Mycology</em> 18, 111-120. The yeast form of <em>Candida albicans</em> is subjected to particle electrophoresis in a variety of media, in order to determine whether the cell surface of the fungus conforms to a simple electrostatic system. It is found that <em>C. albicans</em> behaves essentially as a simple charged colloidal system. Similar measurements were performed with various glass surfaces in order to identify whether electrostatic interactions have any bearing on fungal adhesion. It was found that under all the circumstances studied, the fungi and glass were electronegative; the degree of adhesion was found to be affected by the magnitude of the coulombic repulsion. Significant adhesion still occurred, however, even when the coulombic repulsion was a maximum; this was taken to indicate that the fungal surface possesses other nonelectrostatic forces which are attractive. Both the electrostatic repulsive and the nonelectrostatic (presumably nonpolar) forces are considered to play a role in the adhesion of fungi to each other, to artificial surfaces such as glass, and presumably to other surfaces which occur in living systems.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 2","pages":"Pages 111-120"},"PeriodicalIF":0.0,"publicationDate":"1994-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79848922","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 43
Ribosomal DNA Sequence Analysis Reveals New Species Groupings in the Genus Colletotrichum 核糖体DNA序列分析揭示炭疽菌属的新种群
Experimental Mycology Pub Date : 1994-06-01 DOI: 10.1006/emyc.1994.1014
Christine Sherriff, Mitzi J. Whelan, Gillian M. Arnold, Jean-Francois Lafay, Yves Brygoo, John A. Bailey
{"title":"Ribosomal DNA Sequence Analysis Reveals New Species Groupings in the Genus Colletotrichum","authors":"Christine Sherriff,&nbsp;Mitzi J. Whelan,&nbsp;Gillian M. Arnold,&nbsp;Jean-Francois Lafay,&nbsp;Yves Brygoo,&nbsp;John A. Bailey","doi":"10.1006/emyc.1994.1014","DOIUrl":"10.1006/emyc.1994.1014","url":null,"abstract":"<div><p>Sherriff, C., Whelan, M. J., Arnold, G. M., Lafay, J.-F., Brygoo, Y., and Bailey, J. A. 1994. Ribosomal DNA sequence analysis reveals new species groupings in the genus Colletotrichum. <em>Experimental Mycology</em> 18, 121-138. The relatedness of a range of isolates of <em>Colletotrichum</em> species, selected to represent the major morphological forms of the genus, was studied by comparing their morphology with an analysis of an 886-bp region of their 28S rDNA sequences and ITS-2 regions. rDNA was amplified by PCR. An initial dendrogram for the full sequence was drawn using cluster analysis (UPGMA), with trees then constructed using the neighbor-joining method. The similarity matrices and the resulting dendrogram and trees indicate that the genus can be divided into two groups. One group, consisting of <em>Colletotrichum lindemuthianum, Colletotrichum malvarum, Colletotrichum orbiculare,</em> and <em>Colletotrichum trifolii</em>, was distinct from all the other species. Their rDNA was highly homologous and they had consistent morphological features, including their failure to produce septa during conidial germination, which readily distinguished them from <em>Colletotrichum gloeosporioides</em> and from all other isolates. It is concluded that isolates within this group may represent a single species, which should be referred to as <em>C. orbiculare</em> (Berk. and Mont. (v Arx)), in which distinct host-specific forms exist. Examination of the other group indicates that isolates presently included within <em>C. gloeosporioides</em> represent more than one species. The data also confirmed present day inaccurate identifications; e.g., the causal agent of cowpea anthracnose is not a form of <em>C. lindemuthianum</em> . The distinction between species with falcate conidia, i.e., <em>Colletotrichum capsici, Colletotrichum graminicola, Colletotrichum caudatum,</em> and <em>Colletotrichum truncatum</em>, was verified. The relationships revealed by analysis of a 886-base region between the isolates examined was also evident from selective analysis of domain 2 (206 sites) and ITS-2 (158 sites) sequences, suggesting that further analysis of either of these regions should rapidly extend the understanding of the taxonomy of this important plant pathogenic genus.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 2","pages":"Pages 121-138"},"PeriodicalIF":0.0,"publicationDate":"1994-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1014","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76148157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 169
Preliminary Characterization of Phospholipase A2 in Lagenidium giganteum 巨型蛇舌草磷脂酶A2的初步鉴定
Experimental Mycology Pub Date : 1994-06-01 DOI: 10.1006/emyc.1994.1019
Joanna K MacKichan, Amy R Tuininga, James L Kerwin
{"title":"Preliminary Characterization of Phospholipase A2 in Lagenidium giganteum","authors":"Joanna K MacKichan,&nbsp;Amy R Tuininga,&nbsp;James L Kerwin","doi":"10.1006/emyc.1994.1019","DOIUrl":"10.1006/emyc.1994.1019","url":null,"abstract":"<div><p>MacKichan, J. K., Tuininga, A. R., and Kerwin, J. L. 1994. Preliminary characterization of phospholipase A<sub>2</sub> in <em>Lagenidium giganteum. Experimental Mycology</em> 18, 180-192. Phospholipase A<sub>2</sub> (PLA<sub>2</sub>) hydrolyses the fatty acyl ester bond at the <em>sn</em>-2 position in glycerophospholipids. To better understand its regulatory roles, factors affecting PLA<sub>2</sub> activity in <em>Lagenidium giganteum</em> were investigated: divalent ions; chelators: inhibitors; pH; and substrate concentration. PLA<sub>2</sub> activity of <em>L. giganteum</em> whole cell homogenates was determined using 1-stearoyl-2-[1-<sup>14</sup>C]arachidonoyl phosphatidylcholine as substrate. The divalent cations Ca<sup>2+</sup>, Mg<sup>2+</sup>, and Mn<sup>2+</sup> all enhanced PLA<sup>2</sup> activity, while Co<sup>2+</sup>, Fe<sup>2+</sup>, and Zn<sup>2+</sup> were either slightly inhibitory or without effect. High concentrations of EGTA enhanced activity, low concentrations of the chelators were slightly inhibitory, while high concentrations of EDTA had little effect. EGTA, which has a higher affinity for Ca<sup>2+</sup> and Mn<sup>2+</sup> than Mg<sup>2+</sup>, reduced hydrolysis less than a comparable concentration of EDTA. Two pH optima were found, at both acid (ca. 5.5) and alkaline (ca. 11.5) levels. Four classical inhibitors, nordihydroguaiaretic acid, ellagic acid, gossypol, and 4-bromophenacylbromide, reduced PLA<sub>2</sub> activity by about 80% at 5 m<em>M</em> concentration, 50% with 1 m<em>M</em> inhibitor, and had no effect at 100 μ<em>M</em>. The relatively high levels of these compounds needed to inhibit PLA<sub>2</sub> hydrolysis may have been due to the presence of a cocktail of enzymes, some of which were not susceptible to inhibition. All inhibitors at 1 m<em>M</em> concentration in live cell cultures effectively shut down oosporogenesis, without adverse effects to the mycelia. PLA<sub>2</sub> activity was highest in the late oospore stage of the life cycle, although the enzymes were probably not metabolically active in these stationary cultures. Cultures grown on cholesterol-supplemented defined media had significantly higher levels of PLA<sub>2</sub> activity relative to cultures grown on sterol-free media. The enzyme was found to be associated primarily with microsomal membranes, but there was significant activity in cytosolic fractions. Separation of cell homogenates by column chromatography revealed that there were at least nine enzymes capable of cleaving fatty acids in the <em>sn</em> -2 position of phospholipids.</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 2","pages":"Pages 180-192"},"PeriodicalIF":0.0,"publicationDate":"1994-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1019","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90587277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 7
Preparation of Anti-protein and Anti-mannan Antisera against Fungal Cell Wall by Affinity Chromatography 亲和色谱法制备抗真菌细胞壁蛋白和抗甘露聚糖抗血清
Experimental Mycology Pub Date : 1994-06-01 DOI: 10.1006/emyc.1994.1017
Marı́a Iranzo, Antonio Marcilla, Marı́a Victoria Elorza, Salvador Mormeneo, Rafael Sentandreu
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引用次数: 2
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