Advertisement

Analysis of Angiotensin Metabolism in the Kidney Using Mass Spectrometry

  • Nadja Grobe
  • Khalid M. Elased
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1614)

Abstract

The renin angiotensin system (RAS) is a highly complex enzymatic system consisting of multiple peptide hormones, enzymes, and receptors. A thorough characterization of angiotensin peptide metabolism is crucial for understanding pathological states associated with an imbalanced RAS. Here, we describe two matrix-assisted laser desorption/ionization (MALDI) mass spectrometric (MS) approaches for the assessment of in vitro and in situ RAS enzymatic activities in the kidney using the natural angiotensin peptide substrates. These MS techniques demonstrate high specificity and are superior over conventional spectrophotometric or colorimetric assays since multiple proteolytic cleavage sites can be detected, thus unraveling the complexity of the RAS.

Key words

Mass spectrometry MALDI Imaging Renin-angiotensin system Kidney Enzyme activity Angiotensin metabolism ACE ACE2 NEP 

Notes

Acknowledgment

This work was supported by National Institutes of Health Grants F32 DK-093226 and the Carl W. Gottschalk Research Scholar Grant of the ASN Foundation for Kidney Research to N.G. and R01 HL-093567 to K.M.E.

References

  1. 1.
    Fredline VF, Kovacs EM, Taylor PJ, Johnson AG (1999) Measurement of plasma renin activity with use of HPLC-electrospray-tandem mass spectrometry. Clin Chem 45:659–664PubMedGoogle Scholar
  2. 2.
    Donoghue M, Hsieh F, Baronas E, Godbout K, Gosselin M, Stagliano N, Donovan M, Woolf B, Robison K, Jeyaseelan R, Breitbart RE, Acton S (2000) A novel angiotensin-converting enzyme-related carboxypeptidase (ACE2) converts angiotensin I to angiotensin 1-9. Circ Res 87:E1–E9CrossRefPubMedGoogle Scholar
  3. 3.
    Elased KM, Cool DR, Morris M (2005) Novel mass spectrometric methods for evaluation of plasma angiotensin converting enzyme 1 and renin activity. Hypertension 46:953–959CrossRefPubMedGoogle Scholar
  4. 4.
    Elased KM, Cunha TS, Gurley SB, Coffman TM, Morris M (2006) New mass spectrometric assay for angiotensin-converting enzyme 2 activity. Hypertension 47:1010–1017CrossRefPubMedGoogle Scholar
  5. 5.
    Grobe N, Elased KM, Cool DR, Morris M (2012) Mass spectrometry for the molecular imaging of angiotensin metabolism in kidney. Am J Physiol Endocrinol Metab 302:E1016–E1024CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Grobe N, Di FM, Kashkari N, Chodavarapu H, Somineni HK, Singh R, Elased KM (2015) Functional and molecular evidence for expression of the renin angiotensin system and ADAM17-mediated ACE2 shedding in COS7 cells. Am J Physiol Cell Physiol 308:C767–C777CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Stettner D, Bujak-Gizycka B, Olszanecki R, Rytlewski K, Huras H, Korbut R (2013) Assessment of angiotensin I metabolism in the human placenta using an LC/MS method. Folia Med Cracov 53:31–39PubMedGoogle Scholar
  8. 8.
    Velez JC, Ierardi JL, Bland AM, Morinelli TA, Arthur JM, Raymond JR, Janech MG (2012) Enzymatic processing of angiotensin peptides by human glomerular endothelial cells. Am J Physiol Renal Physiol 302:F1583–F1594CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Pharmacology and Toxicology, Boonshoft School of MedicineWright State UniversityDaytonUSA

Personalised recommendations