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Oya Orun; Pınar Mega Tiber

doi:10.5472/MMJoa.2803.01

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Year 2015, Volume: 28 Issue: 3, 123 - 128, 28.11.2015

Oya Orun Pınar Mega Tiber

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

https://izlik.org/JA69HW27AP

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, Volume: 28 Issue: 3, 123 - 128, 28.11.2015

Oya Orun Pınar Mega Tiber

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

https://izlik.org/JA69HW27AP

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
Authors	Oya Orun This is me Pınar Mega Tiber This is me
Publication Date	November 28, 2015
DOI	https://doi.org/10.5472/MMJoa.2803.01
IZ	https://izlik.org/JA69HW27AP
Published in Issue	Year 2015 Volume: 28 Issue: 3

Cite

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