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Ratio of 5,6,7,8-tetrahydrobiopterin to 7,8-dihydrobiopterin in endothelial cells determines glucose-elicited changes in NO vs. superoxide production by eNOS
Open accessAbstract
5,6,7,8-Tetrahydrobiopterin (BH 4 ) is an essential cofactor of nitric oxide synthases (NOSs). Oxidation of BH 4 , in the setting of diabetes and other chronic vasoinflammatory conditions, can cause cofactor insufficiency and uncoupling of endothelial NOS (eNOS), manifest by a switch from nitric oxide (NO) to superoxide production. Here we tested the hypothesis that eNOS uncoupling is not simply a consequence of BH 4 insufficiency, but rather results from a diminished ratio of BH 4 vs. its catalytically incompetent oxidation product, 7,8-dihydrobiopterin (BH 2 ). In support of this hypothesis, [ 3 H]BH 4 binding studies revealed that BH 4 and BH 2 bind eNOS with equal affinity ( K d ≈ 80 nM) and BH 2 can rapidly and efficiently replace BH 4 in preformed eNOS-BH 4 complexes. Whereas the total biopterin pool of murine endothelial cells (ECs) was unaffected by 48-h exposure to diabetic glucose levels (30 mM), BH 2 levels increased from undetectable to 40% of total biopterin. This BH 2 accumulation was associated with diminished calcium ionophore-evoked NO activity and accelerated superoxide production. Since superoxide production was suppressed by NOS inhibitor treatment, eNOS was implicated as a principal superoxide source. Importantly, BH 4 supplementation of ECs (in low and high glucose-containing media) revealed that calcium ionophore-evoked NO bioactivity correlates with intracellular BH 4 :BH 2 and not absolute intracellular levels of BH 4 . Reciprocally, superoxide production was found to negatively correlate with intracellular BH 4 :BH 2 . Hyperglycemia-associated BH 4 oxidation and NO insufficiency was recapitulated in vivo, in the Zucker diabetic fatty rat model of type 2 diabetes. Together, these findings implicate diminished intracellular BH 4 :BH 2 , rather than BH 4 depletion per se, as the molecular trigger for NO insufficiency in diabetes.
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“iNOS and nNOS show higher affinity toward BH4 than for BH2 (around 10-fold in the absence of l-arginine) (6, 514, 753); however, eNOS shows almost identical binding affinities for BH4 and BH2, and it is thus very sensitive to imbalances in the concentrations of BH4 and BH2 (190).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [190]
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“Thus, rather than the relative amount of BH2, the BH4/BH2 ratio gives a more precise estimation of the endothelial function (190, 192, 974).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [190]
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Cites
Biochemistry and molecular cell biology of diabetic complications
Brownlee, M. (2001) · Nature · doi:10.1038/414813a · Cited by 9,107
Griffith, O. W. (1980) · Analytical Biochemistry · doi:10.1016/0003-2697(80)90139-6 · Cited by 4,628
Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage
Nishikawa, T., Edelstein, D., Du, X. L., Yamagishi, S.-I., Matsumura, T., Kaneda, Y., Yorek, M. A., Beebe, D., Oates, P. J., Hammes, H.-P., et al. (2000) · Nature · doi:10.1038/35008121 · Cited by 4,277
Schächinger, V., Britten, M. B., & Zeiher, A. M. (2000) · Circulation · doi:10.1161/01.cir.101.16.1899 · Cited by 2,589
The reaction of NO with superoxide
Huie, R. E., & Padmaja, S. (1993) · Free Radical Research Communications · doi:10.3109/10715769309145868 · Cited by 2,121
Heitzer, T., Schlinzig, T., Krohn, K., Meinertz, T., & Münzel, T. (2001) · Circulation · doi:10.1161/hc4601.099485 · Cited by 1,961
Landmesser, U., Dikalov, S., Price, S. R., McCann, L., Fukai, T., Holland, S. M., Mitch, W. E., & Harrison, D. G. (2003) · Journal of Clinical Investigation · doi:10.1172/jci200314172 · Cited by 1,660
Landmesser, U., Dikalov, S., Price, S. R., McCann, L. A., Fukai, T., Holland, S. M., Mitch, W. E., & Harrison, D. G. (2003) · Journal of Clinical Investigation · doi:10.1172/jci14172 · Cited by 1,652
The chemistry of peroxynitrite: A product from the reaction of nitric oxide with superoxide
Pryor, W. A., & Squadrito, G. L. (1995) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.1995.268.5.l699 · Cited by 1,511
Superoxide generation by endothelial nitric oxide synthase: The influence of cofactors
Vásquez-Vivar, J., Kalyanaraman, B., Martásek, P., Hogg, N., Masters, B. S. S., Karoui, H., Tordo, P., & Pritchard, K. A. (1998) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.95.16.9220 · Cited by 1,484
Laursen, J. B., Somers, M., Kurz, S., McCann, L., Warnholtz, A., Freeman, B. A., Tarpey, M., Fukai, T., & Harrison, D. G. (2001) · Circulation · doi:10.1161/01.cir.103.9.1282 · Cited by 721
Interactions of Peroxynitrite, Tetrahydrobiopterin, Ascorbic Acid, and Thiols
Kuzkaya, N., Weissmann, N., Harrison, D. G., & Dikalov, S. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m302227200 · Cited by 692
Macrophage Oxidation of L-Arginine to Nitric Oxide, Nitrite, and Nitrate
Tayeh, M. A., & Marletta, M. A. (1989) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)47163-9 · Cited by 687
Zou, M., Shi, C., & Cohen, R. A. (2002) · Journal of Clinical Investigation · doi:10.1172/jci14442 · Cited by 613
Tetrahydrobiopterin restores endothelial function in hypercholesterolemia.
Stroes, E. S. G., Kastelein, J. J. P., Cosentino, F., Erkelens, W. D., Wever, R. M. F., Koomans, H. A., Lüscher, T. F., & Rabelink, T. J. (1997) · Journal of Clinical Investigation · doi:10.1172/jci119131 · Cited by 588
Reduced Biopterin as a Cofactor in the Generation of Nitrogen Oxides by Murine Macrophages
Kwon, N. S., Nathan, C. F., & Stuehr, D. J. (1989) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)47089-0 · Cited by 570
Regulation of Endothelial Nitric Oxide Synthase by Tetrahydrobiopterin in Vascular Disease
Alp, N. J., & Channon, K. M. (2003) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/01.atv.0000110785.96039.f6 · Cited by 536
Zou, M.-H., Shi, C., & Cohen, R. A. (2002) · Journal of Clinical Investigation · doi:10.1172/jci0214442 · Cited by 494
Oxidation of Tetrahydrobiopterin by Peroxynitrite: Implications for Vascular Endothelial Function
Milstien, S., & Katušić, Z. S. (1999) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1999.1422 · Cited by 492
Endothelial Dysfunction in Human Disease
Drexler, H., & Hornig, B. (1999) · Journal of Molecular and Cellular Cardiology · doi:10.1006/jmcc.1998.0843 · Cited by 471
Tetrahydrobiopterin Improves Endothelium-Dependent Vasodilation in Chronic Smokers
Heitzer, T. A., Brockhoff, C. J., Mayer, B., Warnholtz, A., Mollnau, H., Henne, S., Meinertz, T., & Münzel, T. F. (2000) · Circulation Research · doi:10.1161/01.res.86.2.e36 · Cited by 449
Gross, S. S., JAFFE, E. A., Levi, R., & Kilbourn, R. G. (1991) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(91)90965-a · Cited by 389
Self-citationHuang, A., Vita, J. A., Venema, R. C., & Keaney, J. F. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m002248200 · Cited by 365
McMillan, K., Bredt, D. S., Hirsch, D. B., Snyder, S. H., Clark, J. E., & Masters, B. S. S. (1992) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.89.23.11141 · Cited by 365
Heerdt, P. M., Holmes, J. W., Cai, B., Barbone, A., Madigan, J. D., Reiken, S. R., Lee, D. L., Öz, M. C., Marks, A. R., & Burkhoff, D. (2000) · Circulation · doi:10.1161/01.cir.102.22.2713 · Cited by 298
d’Uscio, L. V., Milstien, S., Richardson, D. M., Smith, L., & Katušić, Z. S. (2003) · Circulation Research · doi:10.1161/01.res.0000049166.33035.62 · Cited by 296
VÁSQUEZ-VIVAR, J., MARTÁSEK, P., WHITSETT, J., JOSEPH, J., & KALYANARAMAN, B. (2002) · Biochemical Journal · doi:10.1042/bj3620733 · Cited by 287
LINDSAYWILLIAMS, R. (1988) · Cell · doi:10.1016/0092-8674(88)90536-3 · Cited by 257
Nitric oxide and the endothelium: History and impact on cardiovascular disease
Yetik-Anacak, G., & Catravas, J. D. (2006) · Vascular Pharmacology · doi:10.1016/j.vph.2006.08.002 · Cited by 253
Shinozaki, K., Nishio, Y., Okamura, T., Yoshida, Y., Maegawa, H., Kojima, H., Masada, M., Toda, N., Kikkawa, R., & Kashiwagi, A. (2000) · Circulation Research · doi:10.1161/01.res.87.7.566 · Cited by 242
Srinivasan, S. R., Hatley, M., Bolick, D. T., Palmer, L. A., Edelstein, D., Brownlee, M., & Hedrick, C. C. (2004) · Diabetologia · doi:10.1007/s00125-004-1525-1 · Cited by 216
Brodsky, S. V., Gealekman, O., Chen, J., Zhang, F., Togashi, N., Crabtree, M. J., Gross, S. S., Nasjletti, A., & Goligorsky, M. S. (2003) · Circulation Research · doi:10.1161/01.res.0000111802.09964.ef · Cited by 215
Self-citationHurshman, A. R., Krebs, C., Edmondson, D. E., Huynh, B. H., & Marletta, M. A. (1999) · Biochemistry · doi:10.1021/bi992026c · Cited by 208
Mechanisms for the role of tetrahydrobiopterin in endothelial function and vascular disease
Schmidt, T. S., & Alp, N. J. (2007) · Clinical Science · doi:10.1042/cs20070108 · Cited by 195
Meininger, C. J., MARINOS, R. S., Hatakeyama, K., Martı́nez–Zaguilán, R., Rojas, J. D., Kelly, K. A., & Wu, G. (2000) · Biochemical Journal · doi:10.1042/0264-6021:3490353 · Cited by 186
Nitric oxide dynamics and endothelial dysfunction in type II model of genetic diabetes
Bitar, M. S., Wahid, S., Mustafa, S., Al‐Saleh, E., Dhaunsi, G. S., & Al‐Mulla, F. (2005) · European Journal of Pharmacology · doi:10.1016/j.ejphar.2005.01.014 · Cited by 179
Gori, T., Burstein, J. M., Ahmed, S., Miner, S. E. S., Al-Hesayen, A., Kelly, S., & Parker, J. D. (2001) · Circulation · doi:10.1161/hc3501.095358 · Cited by 178
Nephropathy in Zucker Diabetic Fat Rat Is Associated with Oxidative and Nitrosative Stress
Chander, P. N., Gealekman, O., Brodsky, S. V., Elitok, S., Tojo, A., Crabtree, M. J., Gross, S. S., & Goligorsky, M. S. (2004) · Journal of the American Society of Nephrology · doi:10.1097/01.asn.0000135971.88164.2c · Cited by 174
Self-citationHigashi, Y., Sasaki, S., Nakagawa, K., Fukuda, Y., Matsuura, H., Oshima, T., & Chayama, K. (2002) · American Journal of Hypertension · doi:10.1016/s0895-7061(01)02317-2 · Cited by 173
CAI, S., Khoo, J., & Channon, K. M. (2004) · Cardiovascular Research · doi:10.1016/j.cardiores.2004.10.040 · Cited by 164
Smoking Impairs the Activity of Endothelial Nitric Oxide Synthase in Saphenous Vein
Higman, D. J., Strachan, A. M. J., Buttery, L. D. K., Hicks, R., Springall, D. R., Greenhalgh, R. M., & Powell, J. T. (1996) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/01.atv.16.4.546 · Cited by 155
Antoniades, C., Shirodaria, C., Crabtree, M., Rinze, R., Alp, N., Cunnington, C., Diesch, J., Tousoulis, D., Stefanadis, C., Leeson, P., et al. (2007) · Circulation · doi:10.1161/circulationaha.107.704155 · Cited by 153
Self-citationCharacterization of Bovine Endothelial Nitric Oxide Synthase Expressed inE. coli
Pavel Martasek, E., Liu, Q., Liu, J., Roman, L. J., Gross, S. S., Sessa, W. C., & Masters, B. S. S. (1996) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1996.0238 · Cited by 149
Self-citationRestoration of Endothelium-Dependent Vasodilation After Reperfusion Injury by Tetrahydrobiopterin
Tiefenbacher, C., Chilian, W. M., Mitchell, M., & DeFily, D. V. (1996) · Circulation · doi:10.1161/01.cir.94.6.1423 · Cited by 140
Altered Tetrahydrobiopterin Metabolism in Atherosclerosis
Vasquez-Vivar, J. M., Duquaine, D. C., Whitsett, J., Kalyanaraman, B., & Rajagopalan, S. (2002) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/01.atv.0000029122.79665.d9 · Cited by 123
Vásquez-Vivar, J., Whitsett, J., Martásek, P., Hogg, N., & Kalyanaraman, B. (2001) · Free Radical Biology and Medicine · doi:10.1016/s0891-5849(01)00680-3 · Cited by 112
Paradoxical overexpression and translocation of connexin43 in homocysteine-treated endothelial cells
Li, H., Brodsky, S. V., Kumari, S., Valiūnas, V., Brink, P. R., Kaide, J.-I., Nasjletti, A., & Goligorsky, M. S. (2002) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.01028.2001 · Cited by 95
Self-citationThe Autoxidation of Tetrahydrobiopterin Revisited
Kirsch, M., Korth, H., Stenert, V., Sustmann, R., & de GROOT, H. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m211779200 · Cited by 94
The auto‐oxidation of tetrahydrobiopterin
DAVIS, M. D., Kaufman, S., & Milstien, S. (1988) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1988.tb14004.x · Cited by 94
A simple and sensitive bioassay method for detection of EDRF with RFL-6 rat lung fibroblasts
Ishii, K., Sheng, H., Warner, T. D., Förstermann, U., & Murad, F. (1991) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.1991.261.2.h598 · Cited by 93
Effect of Tetrahydrobiopterin on Endothelial Function in Canine Middle Cerebral Arteries
Tsutsui, M., Milstien, S., & Katušić, Z. S. (1996) · Circulation Research · doi:10.1161/01.res.79.2.336 · Cited by 92
Sawabe, K., Wakasugi, K. O., & Hasegawa, H. (2004) · Journal of Pharmacological Sciences · doi:10.1254/jphs.fp0040280 · Cited by 75
Pieper, G. M., Siebeneich, W., Hilton, G., & Roza, A. M. (1997) · Diabetologia · doi:10.1007/s001250050767 · Cited by 74
Wolff, D. J., Datto, G. A., & Samatovicz, R. A. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)98369-9 · Cited by 70
Setoguchi, S., Hirooka, Y., Eshima, K., Shimokawa, H., & Takeshita, A. (2002) · Journal of Cardiovascular Pharmacology · doi:10.1097/00005344-200203000-00007 · Cited by 57
Measurement of NO and NO Synthase
Weissman, B. A., & Gross, S. S. (1998) · Current Protocols in Neuroscience · doi:10.1002/0471142301.ns0713s05 · Cited by 51
Self-citationEnhanced inflammatory responses in α-tocopherol transfer protein null mice
Schock, B. C., van der Vliet, A., Corbacho, A. M., Leonard, S. W., Finkelstein, E. I., Valacchi, G., Obermueller-Jevic, U. C., Cross, C. E., & Traber, M. G. (2004) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2003.12.009 · Cited by 44
Clinical aspects of reactive oxygen and nitrogen species
Warnholtz, A., Wendt, M., August, M., & Münzel, T. F. (2004) · Biochemical Society Symposia · doi:10.1042/bss0710121 · Cited by 37
Heales, S., & Hyland, K. (1989) · Journal of Chromatography B Biomedical Sciences and Applications · doi:10.1016/s0378-4347(00)82658-4 · Cited by 36
Zanetti, M., Sato, J., Katušić, Z. S., & O’Brien, T. (2000) · Diabetologia · doi:10.1007/s001250050052 · Cited by 34
Britten, M. B., Zeiher, A. M., & chinger, V. S. (2003) · Coronary Artery Disease · doi:10.1097/00019501-200309000-00001 · Cited by 28
Nitric oxide synthesis in retinal photoreceptor cells
Yoshida, A., Pozdnyakov, N., Dang, L., Orselli, S. M., Reddy, V. N., & Sitaramayya, A. (1995) · Visual Neuroscience · doi:10.1017/s0952523800008397 · Cited by 27
Ono, S., Ichikura, T., & Mochizuki, H. (2003) · PubMed · Cited by 10
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Metadata
- Authors
- Crabtree, M. J., Smith, C. L., Lam, G., Goligorsky, M. S., & Gross, S. S.
- Affiliations
- Cornell University (US)
- Venue
- American Journal of Physiology-Heart and Circulatory Physiology
- Publisher
- American Physiological Society
- Published
- 2008-04
- Volume
- 294
- Issue
- 4
- Pages
- H1530-H1540
- ISSN
- 0363-6135 (ISSN-L)
- 1522-1539
- PMC
- PMC2722919
- OpenAlex
- W2148390554
- Cited by
- 213
- References
- 62
- Citations by year
- 2026: 8 · 2025: 8 · 2024: 6 · 2023: 5 · 2022: 5 · 2021: 9 · 2020: 8 · 2019: 5 · 2018: 12 · 2017: 9 · 2016: 20 · 2015: 13 · 2014: 13 · 2013: 20 · 2012: 15
- FWCI
- 12.64
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- American Journal of Physiology-Heart and Circulatory Physiology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Heart, Lung, and Blood Institute (P01HL046403, HL87062, R37HL087062, R01HL080702, HL-46403)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK045462, DK 45462)
- Topics
- Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Keywords
- tetrahydrobiopterin, 7,8-dihydrobiopterin, eNOS uncoupling, nitric oxide production, superoxide production, endothelial cells, hyperglycemia, type 2 diabetes, BH4 supplementation, calcium ionophore, Zucker diabetic fatty rats
- MeSH terms
- Animals, Biopterins, Blood Glucose, Cell Line, Diabetes Mellitus, Type 2, Disease Models, Animal, Enzyme Inhibitors, Glucose, Glutathione, Mitochondria, Nitric Oxide, Oxidation-Reduction, Protein Binding, Rats, Zucker, Superoxides, Time Factors, Tritium, NG-Nitroarginine Methyl Ester, Endothelial Cells, Mice, Rats, Nitric Oxide Synthase Type II, Nitric Oxide Synthase Type III
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