-

Oya Orun; Pınar Mega Tiber

doi:10.5472/MMJoa.2803.01

-

Year 2015, , 123 - 128, 28.11.2015

Oya Orun Pınar Mega Tiber

https://doi.org/10.5472/MMJoa.2803.01

Abstract

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Keywords

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References

1. Amara SG, Kuhar MJ. Neurotransmitter transporters: recent progress. Annu Rev Neurosci 1993; 16: 73-93. doi: 10.1146/ annurev.ne.16.030193.000445
2. Giros B, Caron MG. Molecular characterization of the dopamine transporter. Trends Pharmacol Sci 1993; 14: 43-9. doi:10.1016/0165-6147(93)90029-J
3. Hyman SE, Malenka RC, Nestler EJ. Neural mechanisms of addiction: the role of reward-related learning and memory. Annu Rev Neurosci 2006; 29: 565-98. doi: 10.1146/annurev. neuro.29.051605.113009
4. Yamashita A, Singh SK, Kawate T, et al. Crystal structure of a bacterial homologue of Na+/Cl- dependent neurotransmitter transporters. Nature 2005; 437: 215-23. doi:10.1038/ nature03978.
5. Guptaroy B, Zhang M, Bowton E, et al. A juxtamembrane mutation in the N terminus of the dopamine transporter induces preference for an inward-facing conformation. Mol Pharmacol 2009; 75: 514-24. doi: 10.1124/mol.108.048744.
6. Lin Z, Zhang PW, Zhu X, et al. Phosphatidylinositol 3-kinase, protein kinase C, and MEK1/2 kinase regulation of dopamine transporters (DAT) require N-terminal DAT phosphoacceptor sites. J Biol Chem 2003; 278: 20162-170. doi : 10.1074/jbc. M209584200.
7. Khoshbouei H, Sen N, Guptaroy B, et al. N-terminal phosphorylation of the dopamine transporter is required for amphetamine-induced efflux. PLoS Biology 2004; 2: 0387- 93. doi: 10.1371/journal.pbio.0020078
8. Granas C, Ferrer J, Loland CJ, et al. N-terminal truncation of the dopamine transporter abolishes phorbol ester- and substance P receptor-stimulated phosphorylation without impairing transporter internalization. J Biol Chem 2003; 278: 4990-5000. doi : 10.1074/jbc.M205058200.
9. Förster T. Mechanism of energy transfer. In: Glorkin M and Stotz EH, editors. Vol 22. Comprehensive Biochemistry. Amsterdam:Elsevier, 1967: 61-80.
10. Eriksen J, Bjørn-Yoshimoto WE, Jørgensen TN, et al. Postendocytic sorting of constitutively internalized dopamine transporter in cell lines and dopaminergic neurons. J Biol Chem 2010; 285: 27289-301. doi :10.1074/jbc.M110.131003.
11. Indarte M, Madura JD, Surratt CK. Dopamine transporter comparative molecular modeling and binding site prediction using the LeuTAa leucine transporter as a template. Proteins 2008; 70: 1033-46. doi : 10.1002/prot.21598.
12. Shan J, Javitch JA, Shi L, et al. The substrate-driven transition to an inward-facing conformation in the functional mechanism of the dopamine transporter. PLoS One 6, e16350. doi: 10.1371/journal.pone.0016350
13. Kristensen AS, Andersen J, Jørgensen TN, et al. SLC6 neurotransmitter transporters: structure, function, and regulation. Pharmacol Rev 2011; 63: 585-640. doi: 10.1124/ pr.108.000869.
14. Cervinski MA, Foster JD, Vaughan RA. Psychoactive substrates stimulate dopamine transporter phosphorylation and down-regulation by cocaine-sensitive and protein kinase C-dependent mechanisms. J Biol Chem 2005; 280: 40442-9. doi: 10.1074/jbc.M501969200.
15. Moritz AE, Foster JD, Gorentla BK, et al. Phosphorylation of dopamine transporter serine 7 modulates cocaine analog binding. J Biol Chem 2013; 288: 20-32. doi: 10.1074/jbc. M112.407874

Dopamin taşıyıcıların kokain bağlama bölgesi ile N-ucu arasındaki uzaysal yakınlığının FRET ile belirlenmesi

Year 2015, , 123 - 128, 28.11.2015

Oya Orun Pınar Mega Tiber

https://doi.org/10.5472/MMJoa.2803.01

Abstract

Amaç: Dopamin taşıyıcılar (DAT) dopaminin sinaptik gerialımından
ve böylelikle sinirsel iletinin çabuk sonlanmasından
sorumludur. Bu proteinler alışkanlık yapıcı kokain, amfetamin
gibi maddelerin de etki merkezlerini oluşturur. Proteinin N-ucunda
bulunan Serin 7 ve 12’nin fosforilasyon bakımından kritik amino
asitler olduğu bilinmektedir. Bu çalışmada N-ucu ile kokain
bağlama bölgesi arasındaki uzaklık, yaban tip ve Ser7/12Asp ve
Ser7/12Ala mutantlarında belirlenmiştir.
Gereç ve Yöntem: Sarı-floresan protein (YFP) ve mutasyonlar
DAT’ın N-terminal ucuna 2-aşamalı PCR ile yerleştirilmiştir.
Dopamin geri-alım testleri yapılmış ve konfokal mikroskopta
verici-ağartma yöntemi ile Förster rezonans enerji transferi
(FRET) mesafeleri belirlenmiştir.
Bulgular: Tüm N-ucundan YFP-etiketlenmiş DAT proteinleri
düzgün olarak hücre yüzeyine taşınmıştır. Km değerleri sırasıyla
yaban tip, Ser7/12Ala and Ser7/12/Asp mutantları için 0.76,
0.75 ve 1.24 mM olarak saptanmıştır. FRET etkinliği yaban-tip
DAT için 0.15 olarak bulunmuş ve FRET uzaklığı 68.4 Å olarak
bulunmuştur. Diğer konstraktlarda FRET etkinlikleri negatiftir ve
enerji transferi saptanmamıştır.
Sonuç: Veriler bize N-ucu bölgesinin kokain bağlama bölgesi
ile FRET mesafesinde olduğunu ve kritik serin rezidülerinin
mutasyonlarının (Ser7/12Asp) bu mesafeyi arttırdığını
göstermektedir

Keywords

Dopamin taşıyıcı (DAT), Kokain bağlama, Förster rezonans enerji transferi (FRET), DAT amino terminal, DAT fosforilasyonu

References

1. Amara SG, Kuhar MJ. Neurotransmitter transporters: recent progress. Annu Rev Neurosci 1993; 16: 73-93. doi: 10.1146/ annurev.ne.16.030193.000445
2. Giros B, Caron MG. Molecular characterization of the dopamine transporter. Trends Pharmacol Sci 1993; 14: 43-9. doi:10.1016/0165-6147(93)90029-J
3. Hyman SE, Malenka RC, Nestler EJ. Neural mechanisms of addiction: the role of reward-related learning and memory. Annu Rev Neurosci 2006; 29: 565-98. doi: 10.1146/annurev. neuro.29.051605.113009
4. Yamashita A, Singh SK, Kawate T, et al. Crystal structure of a bacterial homologue of Na+/Cl- dependent neurotransmitter transporters. Nature 2005; 437: 215-23. doi:10.1038/ nature03978.
5. Guptaroy B, Zhang M, Bowton E, et al. A juxtamembrane mutation in the N terminus of the dopamine transporter induces preference for an inward-facing conformation. Mol Pharmacol 2009; 75: 514-24. doi: 10.1124/mol.108.048744.
6. Lin Z, Zhang PW, Zhu X, et al. Phosphatidylinositol 3-kinase, protein kinase C, and MEK1/2 kinase regulation of dopamine transporters (DAT) require N-terminal DAT phosphoacceptor sites. J Biol Chem 2003; 278: 20162-170. doi : 10.1074/jbc. M209584200.
7. Khoshbouei H, Sen N, Guptaroy B, et al. N-terminal phosphorylation of the dopamine transporter is required for amphetamine-induced efflux. PLoS Biology 2004; 2: 0387- 93. doi: 10.1371/journal.pbio.0020078
8. Granas C, Ferrer J, Loland CJ, et al. N-terminal truncation of the dopamine transporter abolishes phorbol ester- and substance P receptor-stimulated phosphorylation without impairing transporter internalization. J Biol Chem 2003; 278: 4990-5000. doi : 10.1074/jbc.M205058200.
9. Förster T. Mechanism of energy transfer. In: Glorkin M and Stotz EH, editors. Vol 22. Comprehensive Biochemistry. Amsterdam:Elsevier, 1967: 61-80.
10. Eriksen J, Bjørn-Yoshimoto WE, Jørgensen TN, et al. Postendocytic sorting of constitutively internalized dopamine transporter in cell lines and dopaminergic neurons. J Biol Chem 2010; 285: 27289-301. doi :10.1074/jbc.M110.131003.
11. Indarte M, Madura JD, Surratt CK. Dopamine transporter comparative molecular modeling and binding site prediction using the LeuTAa leucine transporter as a template. Proteins 2008; 70: 1033-46. doi : 10.1002/prot.21598.
12. Shan J, Javitch JA, Shi L, et al. The substrate-driven transition to an inward-facing conformation in the functional mechanism of the dopamine transporter. PLoS One 6, e16350. doi: 10.1371/journal.pone.0016350
13. Kristensen AS, Andersen J, Jørgensen TN, et al. SLC6 neurotransmitter transporters: structure, function, and regulation. Pharmacol Rev 2011; 63: 585-640. doi: 10.1124/ pr.108.000869.
14. Cervinski MA, Foster JD, Vaughan RA. Psychoactive substrates stimulate dopamine transporter phosphorylation and down-regulation by cocaine-sensitive and protein kinase C-dependent mechanisms. J Biol Chem 2005; 280: 40442-9. doi: 10.1074/jbc.M501969200.
15. Moritz AE, Foster JD, Gorentla BK, et al. Phosphorylation of dopamine transporter serine 7 modulates cocaine analog binding. J Biol Chem 2013; 288: 20-32. doi: 10.1074/jbc. M112.407874

There are 15 citations in total.

Details

Primary Language	EN
Journal Section	Articles
Authors	Oya Orun This is me Pınar Mega Tiber This is me
Publication Date	November 28, 2015
Published in Issue	Year 2015

Cite

APA	Orun, O., & Mega Tiber, P. (2015). -. Marmara Medical Journal, 28(3), 123-128. https://doi.org/10.5472/MMJoa.2803.01
AMA	Orun O, Mega Tiber P. -. Marmara Med J. December 2015;28(3):123-128. doi:10.5472/MMJoa.2803.01
Chicago	Orun, Oya, and Pınar Mega Tiber. “-”. Marmara Medical Journal 28, no. 3 (December 2015): 123-28. https://doi.org/10.5472/MMJoa.2803.01.
EndNote	Orun O, Mega Tiber P (December 1, 2015) -. Marmara Medical Journal 28 3 123–128.
IEEE	O. Orun and P. Mega Tiber, “-”, Marmara Med J, vol. 28, no. 3, pp. 123–128, 2015, doi: 10.5472/MMJoa.2803.01.
ISNAD	Orun, Oya - Mega Tiber, Pınar. “-”. Marmara Medical Journal 28/3 (December 2015), 123-128. https://doi.org/10.5472/MMJoa.2803.01.
JAMA	Orun O, Mega Tiber P. -. Marmara Med J. 2015;28:123–128.
MLA	Orun, Oya and Pınar Mega Tiber. “-”. Marmara Medical Journal, vol. 28, no. 3, 2015, pp. 123-8, doi:10.5472/MMJoa.2803.01.
Vancouver	Orun O, Mega Tiber P. -. Marmara Med J. 2015;28(3):123-8.