Immunogold localization of nodule-specific uricase in developing soybean root nodules

Bosch, K. A. Van den; Newcomb, Eldon H.

doi:10.1007/bf00391217

Cited by 89 publications

(36 citation statements)

References 32 publications

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“…By using a histochemical stain, Triplett (14) found XDH activity in the infected cells. Based on that observation and the presence of uricase activity in the peroxisomes of uninfected cells (8)(9)(10)(11)(12), he suggested that uric acid was the intermediate transported between the two cell types. However, we believe that histochemical staining at the light microscopic level lacks sufficient resolution for XDH to be detected if it is present also in the smaller uninfected cells, given their generally thin layer of cytoplasm and large central vacuole.…”

Section: Discussionmentioning

confidence: 99%

Localization of xanthine dehydrogenase in cowpea root nodules: implications for the interaction between cellular compartments during ureide biogenesis.

Datta

Triplett

Newcomb

1991

Proc. Natl. Acad. Sci. U.S.A.

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Immunocytochemistry was used to assess the location of xanthine dehydrogenase (EC 1.1.1.204) in the infected region of nodules of cowpea (Vigna unguiculata [L.] Walpers cv. Queen Anne Blackeye). Polyclonal antibodies raised against purified cowpea xanthine dehydrogenase were used to localize this enzyme at the electron microscopic level. Sparse nonspecific labeling was observed after treatment of nodule sections with preimmune serum. Although immune serum cross-reacted with the ground cytoplasm of both infected and uninfected cells, significantly more labeling was observed in the uninfected cells. No labeling above background was observed in peroxisomes, mitochondria, proplastids, endoplasmic reticulum, cytoplasmic or peribacteroid membranes, peribacteroid spaces, or bacteroids. The enzyme is soluble and not present in any organelle or membrane. The greater concentration of xanthine dehydrogenase in the uninfected cells suggests that xanthine or a precursor to xanthine, rather than uric acid, is the intermediate that moves from infected to uninfected cells during ureide biogenesis.

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Section: Discussionmentioning

confidence: 99%

Localization of xanthine dehydrogenase in cowpea root nodules: implications for the interaction between cellular compartments during ureide biogenesis.

Datta

Triplett

Newcomb

1991

Proc. Natl. Acad. Sci. U.S.A.

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“…A combination of genetic, biochemical, and physiological methods has shown NodC to be a 46.8-kilodalton (kDa) dimeric, integral outer membrane protein with receptor like structure (12,13), while the 21.8-kDa NodA protein may be a soluble polypeptide (25) involved in synthesis of a diffusible plant growth factor (26). The use of immunocytochemistry to localize nodulation gene products has not previously been reported, although this approach has been employed effectively for the localization of other nodule constituents, including the oxygen-binding protein leghemoglobin (21), the nodule-specific enzyme uricase (18,28,29), and certain components of the peribacteroid and plasma membranes of infected cells (2,3,8 (12,25) and Western immunoblot analyses of nodule tissue and fractionated R. meliloti cells (13,25). For prolonged storage, antibodies were filtersterilized, frozen in liquid nitrogen, and kept at -70°C.…”

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confidence: 99%

Immunogold localization of the NodC and NodA proteins of Rhizobium meliloti

1989

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Monospecific, polyclonal antibodies to the nodC and nodA gene products of Rhizobium meliloti were used in combination with immunogold labeling and transmission electron microscopy to localize the NodC and NodA proteins in cultures of R. meliloti. Both NodC and NodA were detected in the cytoplasm and cell envelope in thin sections of free-living rhizobia treated with luteolin, a known inducer of nod gene expression; however, only NodC was detected on cell surfaces when immunolabeling was performed with intact induced cells. In view of biochemical data characterizing NodC as an outer membrane protein with a large extracellular domain, the pattern of immunolabeling on thin sections suggests that NodC is produced on free cytoplasmic ribosomes prior to assembly in the membrane. The pattern of NodA labeling on thin sections is consistent with biochemical data detecting NodA in both soluble and membrane fractions of NodA-overexpressing strains of R. meliloti.

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“…On the other hand, uricase II and other enzymes of ureide metabolism have been detected in uninfected cells of soybean nodules (4). Specifically, uricase II has been localized within the peroxisomes of these cells (2,22). The enzyme uricase (Urate oxidase, EC 1.7.3.3.)…”

mentioning

confidence: 99%

Purification, cDNA Cloning, and Developmental Expression of the Nodule-Specific Uricase from Phaseolus vulgaris L.

Sánchez

Campos²,

Padilla³

et al. 1987

Plant Physiol.

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Nodule-specific uoricase (uricase I) from Phasolus vulgas L. was purified to homogeneity by chromatographic methods. Purification data indicated that uricas II is approximately 2% of the total soluble protein from mature nodules. Specific antiserum was raised and used to determine the developmental expression and for immunoselection of polysomes. Uricase II was antigenically detected early in nodule development, 2 to 3 days before nitrogen fixation. Uricase-encoding cDNA clones were isolated by hybridizing a nodule-specific pUC9 cDNA library with labeled mRNA from immunoselected polysomes and a 35,000 molecular weight uricase II-encoding cDNA from soybean. An homologous clone (pNF-UR07) was used to assess the expression pattern of the specific transcript during development. Northern-blot analysis indicated that uricase II mRNA is exclusively expressed in nodule tissue.

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Immunogold localization of nodule-specific uricase in developing soybean root nodules

Cited by 89 publications

References 32 publications

Localization of xanthine dehydrogenase in cowpea root nodules: implications for the interaction between cellular compartments during ureide biogenesis.

Localization of xanthine dehydrogenase in cowpea root nodules: implications for the interaction between cellular compartments during ureide biogenesis.

Immunogold localization of the NodC and NodA proteins of Rhizobium meliloti

Purification, cDNA Cloning, and Developmental Expression of the Nodule-Specific Uricase from Phaseolus vulgaris L.

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