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Requirement of argininosuccinate lyase for systemic nitric oxide production

Nature Medicine volume 17, pages 1619–1626 (2011)Cite this article

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Abstract

Nitric oxide (NO) is crucial in diverse physiological and pathological processes. We show that a hypomorphic mouse model of argininosuccinate lyase (encoded by Asl) deficiency has a distinct phenotype of multiorgan dysfunction and NO deficiency. Loss of Asl in both humans and mice leads to reduced NO synthesis, owing to both decreased endogenous arginine synthesis and an impaired ability to use extracellular arginine for NO production. Administration of nitrite, which can be converted into NO in vivo, rescued the manifestations of NO deficiency in hypomorphic Asl mice, and a nitric oxide synthase (NOS)-independent NO donor restored NO-dependent vascular reactivity in humans with ASL deficiency. Mechanistic studies showed that ASL has a structural function in addition to its catalytic activity, by which it contributes to the formation of a multiprotein complex required for NO production. Our data demonstrate a previously unappreciated role for ASL in NOS function and NO homeostasis. Hence, ASL may serve as a target for manipulating NO production in experimental models, as well as for the treatment of NO-related diseases.

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Figure 1: Phenotype of Aslneo/neo mice.
Figure 2: Decreased NO production with ASL deficiency.
Figure 3: Pharmacological rescue with an NO source.
Figure 4: Asl knockdown in piglet endothelial cells decreases nitrite production in response to arginine.
Figure 5: ASL-deficient human subjects and cells do not efficiently generate NO from extracellular arginine.
Figure 6: ASL is required to maintain an NO synthetic complex.

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Acknowledgements

This work was supported by the US National Institutes of Health (DK54450, RR19453, RR00188, GM90310 to B.L., GM07526 and DK081735 to A.E., HL75511 to J.L.A., RR024173 to J.C.M. and DK37175 to W.E.M.). We acknowledge and thank the clinical efforts of M. Mullins, S. Carter, A. Tran, J. Stuff, W. Martinek and the Texas Children's Hospital General Clinical Research Center nursing staff. The mutated human ASL–encoding plasmids were kindly provided by L. Salviati from the Department of Pediatrics, University of Padova, Padova, Italy. We thank the families of the subjects for their kind participation. We greatly appreciate the technical contributions of M. Jiang, E. Munivez, B. Dawson, G. Sule and J. Zhang. We thank L. Castillo for helpful discussions. Figures were produced using Servier Medical Art. A.E., S.C.S.N. and O.A.S. are awardees of the National Urea Cycle Disorder Foundation Research Fellowship. O.A.S. is an awardee of the O'Malley Fellowship of the Urea Cycle Disorders Rare Disease Clinical Research Network. N.S.B. is supported by the American Heart Association-National 0735042N. P.M.C. is an awardee of the Canadian Institute of Health Research clinician-scientist training award.

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Authors and Affiliations

  1. Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA

    Ayelet Erez, Sandesh C S Nagamani, Oleg A Shchelochkov, Muralidhar H Premkumar, Philippe M Campeau, Yuqing Chen, Asad Mian, Terry K Bertin, William E O'Brien & Brendan Lee

  2. Howard Hughes Medical Institute, Houston, Texas, USA

    Yuqing Chen, Terry K Bertin & Brendan Lee

  3. Brown Foundation Institute of Molecular Medicine, University of Texas–Houston Health Science Center, Houston, Texas, USA

    Harsha K Garg, Yaoping Tang & Nathan S Bryan

  4. Division of Cardiology, Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, USA

    Li Li & David G Harrison

  5. Department of Pathology, Texas Children's Hospital, Houston, Texas, USA

    Jennifer O Black

  6. Department of Pediatrics and Center for Human Genetics Research, Vanderbilt University Medical Center, Nashville, Tennessee, USA

    Heng Zeng, Marshall Summar & Judy L Aschner

  7. Department of Medicine, Cardiovascular Research, Baylor College of Medicine, Houston, Texas, USA

    Anilkumar K Reddy

  8. The Methodist DeBakey Heart and Vascular Center, Houston, Texas, USA

    Anilkumar K Reddy

  9. Department of Medicine and Division of Nephrology, Baylor College of Medicine, Houston, Texas, USA

    William E Mitch

  10. Department of Pediatrics/Nutrition, Agricultural Research Service/United States Department of Agriculture, Children′s Nutrition Research Center, Baylor College of Medicine, Houston, Texas, USA

    Juan C Marini

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  1. Ayelet Erez
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Contributions

A.E. generated the mouse models, performed most of the experiments and wrote the manuscript. S.C.S.N. performed the human in vivo and in vitro experiments. O.A.S. conducted the mouse therapy experiments. M.H.P. conducted the immunoprecipitation experiments. P.M.C. helped conducting the mutated ASL experiments. Y.C. generated the mouse models. H.K.G. analyzed the NOx data and helped with the ELISA experiments. L.L. performed western blot experiments. A.M. conducted the human studies. T.K.B. conducted the RT-PCR experiments. J.O.B. performed the histological analysis. H.Z. performed western blot and immunoprecipitation experiments. Y.T. conducted the vascular ring experiments. A.K.R. helped with the blood pressure analysis. M.S. contributed to the conception of the hypothesis. W.E.O. analyzed the biochemical data. D.G.H. contributed to our understanding regarding NOS function. W.E.M. conducted the vessel reactivity assay and helped with assessing creatinine clearance. J.C.M. performed the labeled isotope studies. J.L.A. supervised the experiments performed on the Asl hypomorphic lungs, helped in key analyses and helped in revising of the manuscript. N.S.B. helped with the NOx and NOS data analysis and supervised related experiments. B.L. led and supervised the project through all stages.

Corresponding author

Correspondence to Brendan Lee.

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Erez, A., Nagamani, S., Shchelochkov, O. et al. Requirement of argininosuccinate lyase for systemic nitric oxide production. Nat Med 17, 1619–1626 (2011). https://doi.org/10.1038/nm.2544

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