Selective ligand purification using high-performance affinity beads

Ohtsu, Yoshihiro; Ohba, Reiko; Imamura, Yoshimasa; Kobayashi, Motoo; Hatori, Hidetaka; Zenkoh, Tatsuya; Hatakeyama, Mamoru; Manabe, Takashi; Hino, Motohiro; Yamaguchi, Yuki; Kataoka, Kohsuke; Kawaguchi, Haruma; Watanabe, Hidenobu; Handa, Hiroshi

doi:10.1016/j.ab.2004.10.006

Cited by 27 publications

(19 citation statements)

References 18 publications

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“…NIPSNAP1 is found in neuron but not glial cells such as astrocyte and oligodendrocyte, in particular pyramidal neurons in the cerebral cortex, glutamatergic neurons of the granule cell layer, and ␥-aminobutyric acidergic of the polymorphic layer in the hippocampus, dopaminergic neurons in the midbrain, noradrenergic neurons in the brainstem, Purkinje neurons in the cerebellum, and motor neurons in the spinal cord (9). Furthermore, the NIPSNAP1 protein level is increased during generalized seizures caused by kainite (6), and the NIPSNAP1 mRNA level is reduced in the brain of a mouse model for phenylketonuria, an inborn error of amino acid metabolism caused by phenylalanine hydroxylase deficiency (7). Also, NIPSNAP1 binds molecules bearing maple syrup urine disease (9) and Alzheimer disease (8).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, high-performance affinity latex beads, called SG beads, which are glycidylmethacrylate-covered glycidylmethacrylate-styrene copolymer core beads, have been used successfully to purify various proteins including drug receptors (6,7). SG beads have a number of excellent features, such as low nonspecific protein interaction and high purification efficiency.…”

Section: -Nitrophenylphosphatase Domain and Non-neuronal Snap25-likementioning

confidence: 99%

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Identification of NIPSNAP1 as a Nocistatin-interacting Protein Involving Pain Transmission

Okuda‐Ashitaka

Minami

Tsubouchi

et al. 2012

Journal of Biological Chemistry

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Background: Nocistatin (NST) is involved in pain transmission. Results: 4-Nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1 (NIPSNAP1) is purified by using NST-conjugated beads. The inhibition of tactile allodynia by NST is abolished in NIPSNAP1-deficient mice. Conclusion: NIPSNAP1 interacts with NST and mediates pain modulation by NST. Significance: These results may provide a novel therapeutic target for pathological pain.

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Section: Discussionmentioning

confidence: 99%

Section: -Nitrophenylphosphatase Domain and Non-neuronal Snap25-likementioning

confidence: 99%

Identification of NIPSNAP1 as a Nocistatin-interacting Protein Involving Pain Transmission

Okuda‐Ashitaka

Minami

Tsubouchi

et al. 2012

Journal of Biological Chemistry

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“…Purification of PdTCPP-binding Proteins-SGNEGDEN beads or carboxyl-terminal-succinated modified SGNEGE-DENS beads were prepared as described previously (18). To generate PdTCPP-or TCPP-immobilized beads, PdTCPP or TCPP was incubated with one equivalent of N-hydroxysuccinimide in N,N-dimethylformamide at room temperature for 2 h. Succinated 1 mM PdTCPP or TCPP was incubated with 10 mg of SGNEGDEN beads at room temperature overnight.…”

Section: Methodsmentioning

confidence: 99%

“…LC-MS analysis confirmed that the reaction product was 80% single succinated PdTCPP (data not shown). Succinated PdTCPP was then conjugated to amino-modified SG beads (18). Using PdTCPP-conjugated SG beads, we purified PdTCPP-binding proteins from mitochondrial extracts of HeLa cells.…”

Section: Pdtcpp Accumulates In Mitochondria-to Test Whethermentioning

confidence: 99%

Porphyrin Accumulation in Mitochondria Is Mediated by 2-Oxoglutarate Carrier

et al. 2006

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Heme (Fe-protoporphyrin IX), an endogenous porphyrin derivative, is an essential molecule in living aerobic organisms and plays a role in a variety of physiological processes such as oxygen transport, respiration, and signal transduction. For the biosynthesis of heme or the mitochondrial heme proteins, heme or its biosynthetic precursor porphyrin must be transported into mitochondria from cytosol. The mechanism of porphyrin accumulation in the mitochondrial inner membrane is unclear. In the present study, we analyzed the mechanism of mitochondrial translocation of porphyrin derivatives. We showed that palladium meso-tetra(4-carboxyphenyl)porphyrin (PdTCPP), a phosphorescent porphyrin derivative, accumulated in the mitochondria of several cell lines. Using affinity latex beads, we showed that 2-oxoglutarate carrier (OGC), the mitochondrial transporter of 2-oxoglutarate, bound to PdTCPP, and in vitro PdTCPP inhibited 2-oxoglutarate uptake into mitochondria in a competitive manner (K i ‫؍‬ 15 M). Interestingly, all types of porphyrin derivatives examined in this study competitively inhibited 2-oxoglutarate uptake into mitochondria, including protoporphyrin IX, coproporphyrin III, and hemin. Furthermore, mitochondrial accumulation of porphyrins was inhibited by 2-oxoglutarate or OGC inhibitor. These results suggested that porphyrin accumulation in mitochondria is mediated by OGC and that porphyrins are able to competitively inhibit 2-oxoglutarate uptake into mitochondria. This is the first report of a putative mechanism for accumulation of porphyrins in the mitochondrial inner membrane.Porphyrins consist of a tetrapyrrole ring structure and are most widely and efficiently used in energy metabolism. Heme (Fe-protoporphyrin IX), a porphyrin derivative, is a prosthetic molecule for several hemoproteins, and plays an essential role in various biological processes such as oxygen transport, respiration, and signal transduction. The biosynthesis of heme is a multistep process that starts with the condensation of glycineand succinyl-CoA to form 5-aminolevulinate (1). Heme is ultimately formed in the mitochondrial matrix space following translocation of the heme precursor, co-proporphyrinogen III, which is generated in cytosol, into mitochondria (1). Heme is then utilized in the mitochondrial matrix for the synthesis of a variety of heme proteins such as the cytochromes. Thus, it is necessary for heme biosynthesis and synthesis of the mitochondrial heme proteins that heme and porphyrin precursors must be transported into mitochondria from cytosol. However, the mechanism by which this mitochondrial accumulation occurs is unclear.Some metalloporphyrin derivatives are fluorescence-or phosphorescence-emitting molecules. These molecules undergo a shift from a low energy ground state to a high energy excited state upon exposure to UV light. Fluorescent or phosphorescent light is emitted as the molecules decay from their high energy state back down to the ground state. PdTCPP 3 (Fig. 1A) emits long-lived phosphorescence in respons...

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“…Together these approaches have the potential to powerfully advance drug discovery and development. To this end, we have developed a drug screening system that can seek out hit compounds interacting with target proteins, or functional domains of target proteins, from chemical libraries or mycobacterial extracts [13]. We expect that our affinity nanobead technology will provide high-quality chemogenomic information.…”

Section: Discussionmentioning

confidence: 99%