• 제목/요약/키워드: GABA-T

검색결과 72건 처리시간 0.018초

Detection of Human Taurine Transporter and Production of Monoclonal Antibody

  • An, Hye-Suk;Han, Hee-Chang;Lee, Sun-Min;Park, Taesun;Park, Kun-Koo;Kim, Ha-Won
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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    • pp.102-102
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    • 2001
  • Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

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효소처리에 의한 천도복숭아 음료의 기능성 증진 (Effect of Enzyme Treatment on Functional Properties of Nectarine Beverage)

  • 윤선주;이은탁;조준구;김덕진
    • 한국식품영양과학회지
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    • 제39권9호
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    • pp.1379-1383
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    • 2010
  • 본 연구에서는 효소 처리에 의하여 제조한 천도복숭아 주스에 함유된 총 플라보노이드 함량은 대조구의 경우와 cellulase만을 처리한 처리구의 경우 각각 0.146 mg/mL, 0.148 mg/mL로 유사하였으나 pectinase만을 처리한 처리구와 pectinase와 cellulase를 혼합하여 처리한 처리구의 경우 각각 0.167 mg/mL와 0.171 mg/mL로 대조구보다 높게 나타났다. 항산화작용, 즉 전자공여능(DPPH) 조사한 결과는 효소 처리구들이 대조구의 13.42%보다 높은 28.98% 이상으로 2배 이상의 활성을 나타나 효소처리에 의해 제조된 천도복숭아 주스가 열수추출에 의해 제조된 주스보다 높은 항산화기능이 유지될 수 있음을 유추할 수가 있었다. pH 조건별 아질산염 소거 효과를 비교하여 본 결과 대조구의 경우에서는 아질산염 소거능이 pH 1.2에서 약 79%로 나타났으나 효소 처리구들은 90%를 이상의 결과들을 보였으며, pH 3.0, pH 4.0, pH 6.0에서도 대조구의 경우보다 높게 나타났다. 천도복숭아를 이용하여 제조한 주스의 유리아미노산 분석결과 aspartic acid, serine, alanine, GABA, glutamic acid가 총 유리아미노산 검출양의 86.71%에서 94.14% 범위로 대부분의 유리아미노산을 구성하는 것으로 검출되었으며, ornithine과 taurine 또한 검출이 되었다. 각종 무기원소 함량 조사의 결과는 질소성분(T-N)의 경우 대조구에 비해 효소처리구가 다소 높게 나왔으며, 인성분($P_2O_5$)의 경우는 대조구와 비슷한 결과를 보였으며, 칼슘성분(CaO)의 경우 45 mg/L 이상의 마그네슘성분(MgO)의 경우에서는 85 mg/L 이상의 농도를 보여 역시 높은 함유율을 보였으며, 칼륨성분($K_2O$)의 경우에서는 2,133 mg/L 이상의 농도를 보여 역시 상당히 높은 함유율을 보였다.