Purification of Neuraminidase from Vibrio cholerae

Schramm, Gerhard; Mohr, Elisabeth

doi:10.1038/1831677a0

Cited by 48 publications

(13 citation statements)

References 8 publications

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“…Sialidases from various origins have different molecular weights. Streptomyces sialidases have the lowest value (3.2 x 104) except for V. cholerae enzyme, with a molecular weight of 10I to 2,x 104 reported by Mohr and Schramm (25).…”

Section: Discussionmentioning

confidence: 82%

Purification and Characterization of Streptomyces Sialidases

et al. 1974

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Some strains of Streptomyces produce sialidases. Two sialidases were purified over 1,000-fold from a culture filtrate of two Streptomyces species. They had the same properties in molecular weight, behavior to ions and other reagents, and substrate specificity. They showed very small differences in kinetic properties, pH optima, and heat stability. These Streptomyces sialidases differed markedly from Clostridium perfringens sialidase in molecular weight, p-chloromercuribenzoate sensitivity, and substrate specificity. Approximate molecular weights of the sialidases from Streptomyces and C. perfringens were 32,000 and 57,000, respectively. p-Chloromercuribenzoate (10-I M) caused complete inhibition of C. perfringens sialidase but not of Streptomyces sialidases.Glycosidases called sialidase or neuraminidase (N-acetylneuraminate glycohydrolase [EC 3.2.1.18]) are widely distributed in a variety of microorganisms and animal tissues, and bacterial sialidases have been prepared in a highly purified state (1, 6). However, extensive purification has not been attempted for sialidase from other sources. Sialic acids are constituents of numerous glycoproteins, glycolipids, and oligosaccharides, and in many instances removal of sialic acid by sialidase is accompanied by a loss of biological activity (5,20). Thus, it is thought that a sialidase specifically reacts with these sialyl compounds might be useful in detecting its physiological role.In this communication we describe the purification and characterization of Streptomyces sialidase and compare it with Clostridium perfringens sialidase. MATERIALS AND METHODSOrganisms and growth conditions. Streptomyces strains, to which laboratory numbers (MB395-A5 and MB503-C1) had been given at the Institute of Microbial Chemistry, were studied in detail. Strains MB395-A5 and MB503-C1 were classified as Streptomyces griseus and Streptomyces purpeofuscus, respectively, by M. Hamada in our institute. They were shake-cultured at 130 rpm in a medium containing 1.0% glucose, 1.0% starch, 0.75% peptone, 0.75% meat extract, 0.3% NaCl, 0.1% MgSO4*7H2O, 0.1%

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Section: Discussionmentioning

confidence: 82%

Purification and Characterization of Streptomyces Sialidases

et al. 1974

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“…aliquots of purified neuraminidase diluted in CaBBS to contain 25 units of enzyme per ml. (16). Controls consisted of erythrocytes in CaBBS.…”

Section: Elution Of Parent Strains From Chickenmentioning

confidence: 99%

Studies of Two Kinds of Virus Particles Which Comprise Influenza A2 Virus Strains

Choppin

Tamm

1960

The Journal of Experimental Medicine

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Two kinds of virus particles have been found in varying proportions in influenza A2 strains isolated during the 1957 pandemic. Pure populations of the different particles were obtained, and these substrains were genetically stable on serial passage in the chick embryo. The two virus particles differ markedly in several biological properties though they are antigenically similar. One kind of particle, designated "+," is relatively sensitive to specific antibody, is highly sensitive to inhibition by serum inhibitors and urinary mucoprotein, fails to elute or elutes very slowly from human erythrocytes, and is capable of agglutinating erythrocytes treated extensively with V. cholerae filtrate. The other particle, designated "-," is relatively insensitive to antibodies and urinary mucoprotein, completely insensitive to serum inhibitors, elutes rapidly from erythrocytes, and can agglutinate erythrocytes treated extensively with V. cholerae filtrate. Both "+" and "-" particles destroy virus receptors on urinary mucoprotein. The relative proportions of these two particles determine the characteristics of parent strains in reactions with specific antibody, mucoprotein inhibitors, and erythrocytes. The "+" and "-" particles with several easily identifiable markers are well suited for genetic studies.

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“…Vibrio cholerae neuraminidase was crystallized by Schramm and Mohr (1959) and by Ada and French (1959). The soluble inhibitors were regarded as chemical analogues of the cellular receptors.…”

mentioning

confidence: 99%

“…It also became evident that RDE and the viral enzyme were neuraminidases cleaving specifically the glycosidic linkage joining the N-substituted neuraminic acid to the adjacent sugar residue and that RDE action on red blood cells and soluble inhibitors had the effect described above merely by release of sialic acid residues (see Gottschalk, 1960). Vibrio cholerae neuraminidase was crystallized by Schramm and Mohr (1959) and by Ada and French (1959). The soluble inhibitors were regarded as chemical analogues of the cellular receptors.…”

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confidence: 99%

The Significance of Neuraminic Acid, Serine and Threonine for a Glycoprotein Inhibiting Influenza Virus Haemagglutination

Gottschalk¹

1965

Aust J Exp Biol Med

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Summary.A short review is given of the impact which Sir Macfarlane Burnet's observations, on the influenza virus enzyme and RDE and on the influenza virus haemagglutinin inhibitors, has had on the development of the chemistry of glycoproteins in general and on the elucidation of the molecular structure of neuraminic acid in particular.The structure of the ovine subniaxillary (OSM) glycoprotein, prepared from sheep submaxillary glands, is described and its potency as influenza virus haemagglutinin inhibitor discussed in terms of multiple carbohydrate groups, each terminated by a neuraminic acid residue.Evidence is presented that in OSM (from three-year-old sheep) the predominant linkage of carbohydrate to protein is an O-glycosidic linkage to the hydroxyl group of serine and threonine residues. This linkage is easily split by dilute alkali, most probably by the lnechanisin of ^-elimination.Towards the end of the forties three observations were made at the Walter and Eliza Hall Institute, Melboiime, under the leadership of Sir Macfarlane Burnet, which have done much to renew interest in a nearly forgotten carbohydrate, neuraminic acid, to stimulate research in the long neglected field of mucoproteins and, last, but not least, to elucidaite the basic mechanism by wliich influenza virus particles agglutinate red blood cdls and are adsorbed to host cells. At that time Rurnet and his colleagues were intrigued by Hirst's discovery that the influenza virus particle agglutinates fowl, guinea pig and human eryithrocytes at 4° by being adsorbed to and forming bridges between the red cells. At this temperature, the virus remains attached to the surface of the red cells for more than 15 hours. At 37°, howevei", the virus, once adsorbed at the receptor site of red cells, soon elutes spontaneously leaving behind a red blood cell altered in such a manner so that it can no longer adsorb fresh influenza virus. Hii'St clearly recognized the resemblance of this phenomenon to an enzyme-substrate interaction, the enzyme being possessed by the virus particle and the substrate being embedded in the surface stiucture of the red cell (for review, see Burnet, 1951).

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Purification of Neuraminidase from Vibrio cholerae

Cited by 48 publications

References 8 publications

Purification and Characterization of Streptomyces Sialidases

Purification and Characterization of Streptomyces Sialidases

Studies of Two Kinds of Virus Particles Which Comprise Influenza A2 Virus Strains

The Significance of Neuraminic Acid, Serine and Threonine for a Glycoprotein Inhibiting Influenza Virus Haemagglutination

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