Ferredoxin‐sepharose affinity chromatography for the purification of assimilatory nitrite reductase

Ida, Shoji; Kobayakawa, Kazuya; Morita, Yuhei

doi:10.1016/0014-5793(76)80135-4

Cited by 24 publications

(2 citation statements)

References 15 publications

(16 reference statements)

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“…Nitrite reductase protein was purified in a multi-step procedure from 2 kg of green leaves of fourweek-old mustard plants which were supplied with 15 mM KNO3. In developing the purification procedure, works published by Vega and Kamin (1977), Ida et al (1976) and Ida and Mikami (1986) served as a basis.…”

Section: Methodsmentioning

confidence: 99%

“…Ferredoxin-Sepharose 4B was prepared as described by Ida et al (1976) with some modifications: Cyanogen-bromide-activated Sepharose 4B (Pharmacia) was soaked in 1 mM HC1 (15 min) and then washed with the same solution. The gel was then washed with coupling buffer (0.1 M NaHCO3, 0.5 M NaC1, pH =8.15) and immediately mixed with 20 mg of ferredoxin, dissolved in coupling buffer (spinach-ferredoxin was from Sigma chemicals, Deisenhofen, FRG).…”

Section: Methodsmentioning

confidence: 99%

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Appearance of nitrite-reductase mRNA in mustard seedling cotyledons is regulated by phytochrome

Schuster

Mohr

1990

Planta

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Nitrate reductase (NR, EC 1.6.6.1) and nitrite reductase (NIR, EC 1.7.7.1) are the key enzymes of nitrate reduction. It is well established that the appearance of these enzymes is "induced" by nitrate, and it is generally believed that NR is cytosolic while NIR is plastidic. In mustard (Sinapis alba L.) cotyledons we observed two isoforms of NIR (NIR1 and NIR2) using a chromato-focusing technique. Only one of them (NIR2) disappeared when the plastids were damaged by photooxidation in the presence of Norflurazon. It is concluded that NIR2 is plastidic while NIR1 is extraplastidic and not affected by photooxidation of the plastids. Both isoforms appear to have the same molecular weight (60 kilodaltons, kDa). Two distinct translation products which could be immunoprecipitated with NIR antiserum produced against total NIR from mustard were observed which differed slightly in molecular weight (60 versus 63 kDa). The 63-kDa polypeptide was considered to be the precursor of NIR2. While synthesis of NIR protein depended largely on nitrate, the levels of in-vitro-translatable NIR mRNAs were found to be either independent of nitrate and light (NIR1) or controlled by phytochrome only (NIR2). It appears that phytochrome strongly stimulates the level of mRNA while significant enzyme synthesis (NIR2) takes place only in the presence of relatively large amounts of nitrate. Since an increased enzyme level was strictly correlated with an increase of immunoresponsive NIR protein it is improbable that activation of a precursor plays a role. Rather, it is concluded that, in situ, nitrate controls translation.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Appearance of nitrite-reductase mRNA in mustard seedling cotyledons is regulated by phytochrome

Schuster

Mohr

1990

Planta

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The use of selective adsorbents in capillary electrophoresis-mass spectrometry for analyte preconcentration and microreactions: A powerful three-dimensional tool for multiple chemical and biological applications

Guzman¹,

Stubbs²

2001

Electrophoresis

113

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Much attention has recently been directed to the development and application of online sample preconcentration and microreactions in capillary electrophoresis using selective adsorbents based on chemical or biological specificity. The basic principle involves two interacting chemical or biological systems with high selectivity and affinity for each other. These molecular interactions in nature usually involve noncovalent and reversible chemical processes. Properly bound to a solid support, an "affinity ligand" can selectively adsorb a "target analyte" found in a simple or complex mixture at a wide range of concentrations. As a result, the isolated analyte is enriched and highly purified. When this affinity technique, allowing noncovalent chemical interactions and biochemical reactions to occur, is coupled on-line to high-resolution capillary electrophoresis and mass spectrometry, a powerful tool of chemical and biological information is created. This paper describes the concept of biological recognition and affinity interaction on-line with high-resolution separation, the fabrication of an "analyte concentrator-microreactor", optimization conditions of adsorption and desorption, the coupling to mass spectrometry, and various applications of clinical and pharmaceutical interest.

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ESR of Iron Proteins

Smith¹,

Pilbrow²

1980

Biological Magnetic Resonance

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Ferredoxin‐sepharose affinity chromatography for the purification of assimilatory nitrite reductase

Cited by 24 publications

References 15 publications

Appearance of nitrite-reductase mRNA in mustard seedling cotyledons is regulated by phytochrome

Appearance of nitrite-reductase mRNA in mustard seedling cotyledons is regulated by phytochrome

The use of selective adsorbents in capillary electrophoresis-mass spectrometry for analyte preconcentration and microreactions: A powerful three-dimensional tool for multiple chemical and biological applications

ESR of Iron Proteins

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