Phosphatase activity as a criterion for differentiation of oral mycoplasmas

Shibata, Kenichiro; Totsuka, Masayoshi; Watanabe, Takehiko

doi:10.1128/jcm.23.5.970-972.1986

Cited by 13 publications

(9 citation statements)

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“…ACPs are found in mycoplasmas (16,17), but they have never been purified and characterized, probably because of uncertainty regarding the physiological importance. In this study, an ACP was purified from the membranes of M. fermentans.…”

Section: Discussionmentioning

confidence: 99%

“…This is mainly due to uncertainty regarding their endogenous targets, physiological substrates. Previously, we demonstrated that ACP activity is present in some human-derived mycoplasmas and was higher in Mycoplasma fermentans, Mycoplasma faucium and Mycoplasma buccale than in the other mycoplasmas tested (17). Recently, a strain of M. fermentans has been implicated as a cofactor that would explain various puzzling features of infection by the human immunodeficiency virus (12,14).…”

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confidence: 95%

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Purification and Characterization of an Acid Phosphatase from Mycoplasma fermentans

Noda

Shibata

Sawa

et al. 1994

Microbiology and Immunology

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An acid phosphatase associated with the cell membranes of Mycoplasma fermentans was released from the membranes with Triton X-100, then purified by ion-exchange chromatography on DEAE-Sephacel and CM-Sepharose, followed by affinity chromatography on Con A-Sepharose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme revealed a single band with a molecular mass of 31.2 kilodaltons. The enzyme activity toward p-nitrophenyl phosphate was enhanced remarkably by Cu2+, Co2+ and Mg2+, but the activity was not inhibited by EDTA. The enzyme dephosphorylated O-phospho-L-tyrosine as well as p-nitrophenyl phosphate, but not O-phospho-L-threonine, O-phospho-L-serine, glucose-l-phosphate, phosphoryl choline and adenosine triphosphate. The level of the O-phospho-L-tyrosine phosphatase activity was the highest in Mycoplasma faucium and the second highest in Mycoplasma fermentans of all tested human mycoplasmas.Key words: Mycoplasma fermentans, Acid phosphatase Phosphohydrolases are enzymes that catalyze the hydrolysis of phosphate esters. They play a vital role in regulating the physiological levels of inorganic phosphate and phosphorylated metabolites, as well as the activity of some phosphorylated enzymes (10).Although acid phosphatases (ACPs) are widely distributed in bacteria as well as eukaryotic cells, very little is known about their physiological involvement. This is mainly due to uncertainty regarding their endogenous targets, physiological substrates. Previously, we demonstrated that ACP activity is present in some human-derived mycoplasmas and was higher in Mycoplasma fermentans, Mycoplasma faucium and Mycoplasma buccale than in the other mycoplasmas tested (17). Recently, a strain of M. fermentans has been implicated as a cofactor that would explain various puzzling features of infection by the human immunodeficiency virus (12,14).In this study, we therefore focused upon the ACP of M. fermentans, and purified and characterized the enzyme as the first step in gaining insight into its physiological involvement and pathological roles in human diseases. Materials and MethodsChemicals. p-Nitrophenyl phosphate (PN PP) was purchased from Wako Pure Chemical Industries, Ltd., Tokyo. O-Phospho-L-tyrosine (P-Tyr), O-phospho-L-threonine, O-phospho-L-serine, glucose-1-phosphate, phosphoryl choline and adenosine triphosphate were purchased from Sigma Chemicals Co., Ltd., St. Louis, Mo., U.S.A. Other chemicals were also obtained from commercial sources and were of analytical or reagent grade.Organisms and culture conditions. M. fermentans incognitis was kindly provided by Dr.

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Section: Discussionmentioning

confidence: 99%

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confidence: 95%

Purification and Characterization of an Acid Phosphatase from Mycoplasma fermentans

Noda

Shibata

Sawa

et al. 1994

Microbiology and Immunology

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“…More recent results using a nested PCR reaction detected the organism in the synovial fluid of 23/25 (92%) of rheumatoid arthritis patients, and in 6/7 patients with other inflammatory arthritides, but not in 10 osteoarthritis patients, who do not have an inflammatory cell infiltrate. 29 Noda et al and Shibata et al 30,31 found that M. fermentans also had an acid phosphatase which would hydrolyse p-nitrophenol phosphate; its pH optimum was 5Á0.…”

Section: M M U N O L O G Y O R I G I N a L A R T I C L Ementioning

confidence: 98%

“…However at pH 5Á0 the mycoplasma phosphatase activity against AMP remained almost the same, but was now 11% inhibited by b-glycerophosphate, and the mycoplasma showed some ability to hydrolyse p-nitrophenyl phosphate, indicating that acid phosphatase was present as previously reported. 30,31 It is possible that the only 60% inhibition of the mycoplasmal 5 0 N by AMPCP was a result of some residual acid phosphatase activity against AMP, which may well be a preferred substrate compared to b-glycerophosphate or p-nitrophenyl phosphate.…”

Section: The Nature Of the Induced Enzymementioning

confidence: 99%

The importance of B‐cells and ecto‐5′nucleotidase in Mycoplasma fermentans infection and the relevance to rheumatoid arthritis

2007

Immunology

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Summary The aim of this work was to discover if Mycoplasma fermentans, which is known to infect B cells, could be the cause of the raised ecto‐5′‐nucleotidase observed in the synovial fluid of rheumatoid arthritis patients. The ecto‐5′‐nucleotidase activity in the patients' serum has been shown to correlate with the erythrocyte sedimentation rate and DNA from the mycoplasma has been found in the synovial fluid. B lymphoblastoid cell lines were exposed to 16 strains of Mycoplasma fermentans and their ecto‐5′‐nucleotidase, CD73, was measured both biochemically and by mouse antibodies to human ecto 5′‐nucleotidase using the fluorescence activated cell sorter. The type strain, PG 18, did not grow with the B cells. Some of the mycoplasma strains (9/15) increased the cellular ecto‐5′‐nucleotidase activity from twice to 17 fold, and usually showed 5′‐nucleotidase activity themselves. At least one strain, M106, induced human 5′‐nucleotidase on the normally 5′‐nucleotidase negative Daudi and Raji Burkitt's lymphoma cell lines, and increased sevenfold the 5′‐nucleotidase on the monocyte/macrophage cell line THP‐1. Growing the cells in aged medium increased the level of mycoplasma infection. This mycoplasma‐induced enzyme showed a conformational change and an increase in activity with a glycosylation change involving mannose groups. The other group of strains, mostly of respiratory or cell culture origin, usually did not have any 5′‐nucleotidase of their own and decreased the B‐cell enzyme activity by about half. Electron microscopy and flow cytometry showed that the strain M106 was filamentous and could be found inside the B‐cells. The 5′‐nucleotidase‐inducing strains of M. fermentans may be important in the aetiology of rheumatoid arthritis.

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“…We investigated whether Raytide phosphatase activity existed in other mycoplasmas. It was found that mycoplasmas possessing relatively higher levels of ACP activity (15) had higher levels of specific activity ( Table 2). Although ACPs are widely distributed in bacteria, very little is known about their physiological and pathological involvements.…”

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Acid phosphatase purified from Mycoplasma fermentans has protein tyrosine phosphatase-like activity

et al. 1994

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Phosphatase activity as a criterion for differentiation of oral mycoplasmas

Cited by 13 publications

References 9 publications

Purification and Characterization of an Acid Phosphatase from Mycoplasma fermentans

Purification and Characterization of an Acid Phosphatase from Mycoplasma fermentans

The importance of B‐cells and ecto‐5′nucleotidase in Mycoplasma fermentans infection and the relevance to rheumatoid arthritis

Acid phosphatase purified from Mycoplasma fermentans has protein tyrosine phosphatase-like activity

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