Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Expression and Role of the System L Amino Acid Transporter in FOB Human Osteoblast Cells

사람 골모세포 FOB에서 아미노산 수송계 L의 발현 및 역할

  • Kim, Chang-Hyun (Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Park, Joo-Cheol (Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Kim, Do Kyung (Oral Biology Research Institute, College of Dentistry, Chosun University)
  • 김창현 (조선대학교 치과대학 구강생물학연구소) ;
  • 박주철 (조선대학교 치과대학 구강생물학연구소) ;
  • 김도경 (조선대학교 치과대학 구강생물학연구소)
  • Published : 2005.11.01
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Abstract

Amino acid transporters play an important role in supplying nutrition to normal and cancer cells for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the $Na^+$-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-tyre amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In the present study, we have examined the expression and functional characterization of system L amino acid transporters in FOB human osteoblast cells. RT-PCR and western blot analysis have revealed that the FOB cells expressed LAT1, LAT2 together with their associating protein 4F2hc. The uptakes of $[^{14}C]_L$-leucine by FOB cells are $Na^+$-independent and almost completely inhibited by system L amino acid transporter selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). These results suggest that the transport of neutral amino acids including several essential amino acids for cellular nutrition into the FOB human osteoblast cells is mediated by system L amino acid transporters.

사람의 정상 골모세포 FOB에서 아미노산 수송계 L의 발현 및 이들 수송계 L을 통한 아미노산 수송특성을 밝히기 위하여, FOB 세포에서 RT-PCR, western blot 분석 및 아미노산 uptake 실험 등을 수행하여 다음과 같은 결과들을 얻었다. FOB 세포에서 아미노산 수송계 L의 두 아형인 LAT1, LAT2및 그들의 보조인자 4F2hc의 발현을 확인할 수 있었다. FOB 세포에서 $_{L}-leucine$의 수송은 $Na^+$-비의존적이었다. FOB 세포에서 $_{L}-leucine$의 수송은 아미노산 수송계 L의 선택적 억제제인 BCH에 의해 완전히 차단되었다. FOB 세포에서 여러 아미노산들에 의한 L-leucine수송억제 실험결과는 Xenopus oocyte에서 시행되어 보고되어진 LAT1과 LAT2 수송억제 실험 결과의 특성을 모두 포함하였다. 본 연구의 결과로 사람의 정상 골모세포주인 FOB에서 는 세포성장 및 증식을 위한 중성 아미노산의 수송에 아미노산 수송계 L의 두 아형인 LAT1과 LAT2가 중요한 역할을 하고 있음을 확인할 수 있었다.

Keywords

References

  1. Christensen HN. 1990. Role of amino acid transport and countertransport in nutrition and metabolism. Physiol Rev 70: 43-77 https://doi.org/10.1152/physrev.1990.70.1.43
  2. Gomes P, Soares-da-Silva P. 1999. L-DOPA transport properties in an immortalised cell line of rat capillary cerebral endothelial cells, RBE 4. Brain Res 829: 143- 150 https://doi.org/10.1016/S0006-8993(99)01387-6
  3. Lakshmanan M, Goncalves E, Lessly G, Foti D, Robbins J. 1990. The transport of thyroxine into mouse neurobla-stoma cells, NB41A3: the effect of L-system amino acids. Endocrinology 126: 3245-3250 https://doi.org/10.1210/endo-126-6-3245
  4. Kanai Y, Segawa H, Miyamoto K, Uchino H, Takeda E, Endou H. 1998. Expression cloning and characterization of a transporter for large neutral amino acids activated by the heavy chain of 4F2 antigen (CD98). J Biol Chem 273: 23629-23632 https://doi.org/10.1074/jbc.273.37.23629
  5. Yanagida O, Kanai Y, Chairoungdua A, Kim DK, Segawa H, Nii T, Cha SH, Matsuo H, Fukushima J, Fukasawa Y, Tani Y, Taketani Y, Uchino H, Kim JY, Inatomi J, Okayasu I, Miyamoto K, Takeda E, Goya T, Endou H. 2001. Human L -type amino acid transporter 1 (LAT1): characterization of function and expression in tumor cell lines. Biochim Biophys Acta 1514: 291 -302 https://doi.org/10.1016/S0005-2736(01)00384-4
  6. Uchino H, Kanai Y, Kim DK, Wempe MF, Chairoungdua A, Morimoto E, Anders MW, Endou H. 2002. Transport of amino acid-related compounds mediated by L-type amino acid transporter 1 (LAT1): insights into the mechanisms of substrate recognition. Mol Pharmacol 61: 729-737 https://doi.org/10.1124/mol.61.4.729
  7. Mastroberardino L, Spindler B, Pfeiffer R, Skelly PJ, Loffing J, Shoemaker CB, Verrey F. 1998. Amino-acid transport by heterodimers of 4F2hc/CD98 and members of a permease family. Nature 395: 288-291 https://doi.org/10.1038/26246
  8. Pfeiffer R, Spindler B, Loffing J, Skelly PJ, Shoemaker CB, Verrey F. 1998. Functional heterodimeric amino acid transporters lacking cysteine residues involved in disulfide bond. FEBS Lett 439: 157-162 https://doi.org/10.1016/S0014-5793(98)01359-3
  9. Mannion BA, Kolesnikova TV, Lin SH, Wang S, Thompson NL, Hemler ME. 1998. The light chain of CD98 is identified as E16/TA1 protein. J Biol Chem 273: 33127-33129 https://doi.org/10.1074/jbc.273.50.33127
  10. Nakamura E, Sato M, Yang H, Miyagawa F, Harasaki M, Tomita K, Matsuoka S, Noma A, Iwai K, Minato N. 1999. 4F2 (CD98) heavy chain is associated covalently with an amino acid transporter and controls intracellular trafficking and membrane topology of 4F2 heterodimer. J Biol Chem 274: 3009-3016 https://doi.org/10.1074/jbc.274.5.3009
  11. Sang J, Lim YP, Panzica M, Finch P, Thompson NL. 1995. TA1, a highly conserved oncofetal complementary DNA from rat hepatoma, encodes an integral membrane protein associated with liver development, carcinogenesis, and cell activation. Cancer Res 55: 1152- 1159
  12. Wolf DA, Wang S, Panzica MA, Bassily NH, Thompson NL. 1996. Expression of a highly conserved oncofetal gene, TA1/E16, in human colon carcinoma and other primary cancers: homology to Schistosoma mansoni amino acid permease and Caenorhabditis elegans gene products. Cancer Res 56: 5012-5022
  13. Verrey F, Meier C, Rossier G, Kuhn LC. 2000. Glyco-protein-associated amino acid exchangers: broadening the range of transport specificity. Pflugers Arch 440: 503-512 https://doi.org/10.1007/s004240000274
  14. Pineda M, Fernandez E, Torrents D, Estevez R, Lopez C, Camps M, Lloberas J, Zorzano A, Palacin M. 1999. Identification of a membrane protein, LAT-2, that co-expressed with 4F2 heavy chain, an L-type amino acid transport activity with broad specificity for small large zwitterionic amino acids. J Biol Chem 274: 19738-19744 https://doi.org/10.1074/jbc.274.28.19738
  15. Segawa H, Fukasawa Y, Miyamoto K, Takeda E, Endou H, Kanai Y. 1999. Identification and functional characterization of a $Na^{+}$-independent neutral amino acid transporter with broad substrate selectivity. J Biol Chem 274: 19745-19751 https://doi.org/10.1074/jbc.274.28.19745
  16. Rossier G, Meier C, Bauch C, Summa V, Sordat B, Verrey F, Kuhn LC. 1999. LAT2, a new basolateral 4F2hc/CD98-associated amino acid transporter of kidney and intestine. J Biol Chem 274: 34948-34954 https://doi.org/10.1074/jbc.274.49.34948
  17. Utsunomiya-Tate N, Endou H, Kanai Y. 1996. Cloning and functional characterization of a system ASC-like $Na^{+}$-dependent neutral amino acid tansporter. J Biol Chem 271: 14883-14890 https://doi.org/10.1074/jbc.271.25.14883
  18. Utsunomiya-Tate N, Endou H, Kanai Y. 1997. Tissue specific variants of glutamate transporter GLT-1. FEBS Lett 416: 312-316 https://doi.org/10.1016/S0014-5793(97)01232-5
  19. Kim DK, Kanai Y, Choi HW, Tangtrongsup S, Chair-oungdua A, Babu E, Tachampa K, Anzai N, Iribe Y, Endou H. 2002. Characterization of the system L amino acid transporter in T24 human bladder carcinoma cells. Biochim Biophys Acta 1565: 112-121 https://doi.org/10.1016/S0005-2736(02)00516-3
  20. Kim DK, Yoon JH, Jeon JE, Lee SH. 2004. Expression and functional characterization of neutral amino acid transporter in Hep2 human head and neck squamous cell carcinoma. Int J Oral Biol 29: 51-58
  21. Yoon JH, Kim IJ, Kim H, Kim HJ, Jeong MJ, Ahn SG, Kim SA, Lee CH, Choi BK, Kim JK, Jung KY, Lee SU, Kanai Y, Endou H, Kim DK. 2004. Amino acid transport system L is differently expressed in human normal oral keratinocytes and humal oral cancer cells. Cancer Lett 222: 237-245 https://doi.org/10.1016/j.canlet.2004.09.040
  22. Kim DK, Kim IJ, Hwang SA, Kook JH, Lee MC, Shin BA, Bae CS, Yoon SH, Ahn SG, Kim SA, Kanai Y, Endou H, Kim JK. 2004. System L-amino acid transporters are differently expressed in rat astrocyte and C6 glioma cells. Neuroscience Res 50: 437-446 https://doi.org/10.1016/j.neures.2004.08.003