Recombinant Proteases of the Renin-Angiotensin System (RAS)

The Renin-Angiotensin System (RAS) plays a critical role in maintaining blood pressure homeostasis as well as fluid and salt balance in mammals. Production of angiotensins from angiotensinogen requires the participation and coordination of many proteases in different pathways (Figure 1).1, 2, 3, 4, 5, 6, 7 R&D Systems now offers RAS-related recombinant proteases that can be used in studies of their structure-function relationship and in development of their activators and inhibitors.

R&D Systems offers recombinant proteases of RAS
Protein Products Catalog # Form
Renin Human Renin 4090-AS Pro

Activator: Trypsin 3 (Catalog # 3714-SE)
Assay Substrate: RE(EDANS)IHPFHLVIHTK (DABCYL)R
Angiotensin I-
converting enzyme		 Human ACE
Mouse ACE		 929-ZN
1513-ZN		 Ecto
Ecto

Assay Substrate: Mca-RPPGFSAFK(Dnp)-OH
(Catalog # ES005)
Angiotensin I-
converting enzyme 2		 Human ACE2
Mouse ACE2		 933-ZN
3437-ZN		 Ecto
Ecto

Assay Substrate: Mca-YVADAPK(Dnp)-OH
(Catalog # ES007)
Chymase Human CMA1 4099-SE Pro

Activator: Cathepsin C (Catalog # 2336-CY)
Assay Substrate: Suc-AAPF-AMC
Cathespin A Human CTSA
Mouse CTSA		 1049-SE
9789-SE		 Pro
Pro

Activator: Cathepsin L (Catalog # 952-CY)
Assay Substrate: Mca-RPPGFSAFK(Dnp)-OH
(Catalog # ES005)
Aminopeptidase A Human APA 2499-ZN Mature

Assay Substrate: E-AMC
Aminopeptidase N Human APN
Mouse APN		 3815-ZN
2335-ZN		 Ecto
Ecto

Assay Substrate: A-AMC
Neprilysin Human NEP
Mouse NEP		 1182-ZNC
1126-ZNC		 Ecto
Ecto

Assay Substrate: Mca-RPPGFSAFK(Dnp)-OH
(Catalog # ES005)
Thimet oligopeptidase Human THOP1 3439-ZN Full Length

Assay Substrate: Mca-PLGPK(Dnp)-OH
Neurolysin Human NLN 3814-ZN Full Length

Assay Substrate: Mca-PLGPK(Dnp)-OH

Proteases of RAS
Figure 1. A cascade of proteolytic reactions in RAS results in the generation of different angiotensin (Ang) peptides.
View Larger Image 		Figure 1. A cascade of proteolytic reactions in RAS results in the generation of different angiotensin (Ang) peptides. Renin cleaves the precursor protein, angiotensinogen, releasing the inactive peptide, angiotensin I. The Carboxypeptidases ACE, ACE2, CMA1, CTSA and CPA3, the aminopeptidases APA and APN, and the endopeptidases CTSG, NEP, THOP1 and NLN participate in various pathways to generate Ang peptides with diverse functions. For example, Ang II and III act as vasoconstrictors whereas Ang1-7 acts as a vasodilator. Active Ang peptides function through their respective receptors, and the same Ang peptides may have different effects through different receptors (not shown).

Optimization of Chymase Activation
The conversion of prochymase into active chymase is mediated by recombinant mouse Cathepsin C (Catalog # 2336-CY) in the presence of heparin.
View Larger Image 		Figure 2. The conversion of prochymase into active chymase is mediated by Recombinant Mouse Cathepsin C (Catalog # 2336-CY) in the presence of heparin. The optimal concentrations of heparin and Cathepsin C were obtained by fixing Cathep­sin C (10 µg/mL, top graph) or heparin (0.05 mg/mL, bottom graph).

References

  1. Kramkowski, K. et al. (2006) J. Physiol. Pharmacol.57:529.
  2. Thomas, M.C. & C. Tikellis (2005) Curr. Enzyme Inhibition 1:51.
  3. Eriksson, U. et al. (2002) Curr. Biol. 12:R745.
  4. Turner, A.J. & N.M. Hooper (2002) Trends Pharmacol. Sci. 23:177.
  5. Jackman, H.L. et al. (2002) Hypertension 39:976.
  6. Lunderquist, A. et al. (2004) J. Biol. Chem. 279:32339.
  7. Shrimpton, C.N. et al. (2002) Endocr. Rev. 23:647.