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Related Experiment Videos

Rat intestinal angiotensin-converting enzyme: purification, properties, expression, and function.

R H Erickson1, Y Suzuki, A Sedlmayer

  • 1Gastrointestinal Research Laboratory, Department of Veterans Affairs Medical Center, San Francisco, California 94121.

The American Journal of Physiology
|October 1, 1992
PubMed
Summary
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Researchers purified rat intestinal angiotensin-converting enzyme (ACE), a metalloglycoprotein crucial for dietary peptide digestion. ACE expression is regulated by mRNA levels and concentrated in the upper small intestine.

Area of Science:

  • Biochemistry
  • Gastroenterology
  • Molecular Biology

Background:

  • Angiotensin-converting enzyme (ACE) plays a role in peptide metabolism.
  • Understanding ACE's function in the intestine is important for digestive physiology.

Purpose of the Study:

  • To purify and characterize ACE from rat intestinal mucosa.
  • To investigate the distribution and regulation of ACE in the small intestine.
  • To determine ACE's role in dietary peptide assimilation.

Main Methods:

  • Purification of ACE using affinity chromatography (lisinopril-Sepharose) and gel filtration.
  • Homogeneity and molecular weight determination via SDS-PAGE.
  • Enzyme kinetics and inhibitor studies.
  • Analysis of ACE activity, mRNA, and protein distribution along the rat small intestine.

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Main Results:

  • ACE was purified 4,500-fold to homogeneity (160,000 MW), revealing it as a metalloglycoprotein with 12% N-linked carbohydrate.
  • Optimal activity was observed with substrates having proline at the COOH-terminus.
  • ACE activity, mRNA, and protein were highest in the proximal-middle small intestine, decreasing distally.
  • In vivo jejunal perfusion confirmed ACE's role as a dipeptidyl carboxypeptidase in dietary peptide digestion.

Conclusions:

  • Rat intestinal ACE is a metalloglycoprotein primarily functioning in the proximal small intestine.
  • ACE expression appears to be regulated at the mRNA level.
  • ACE is integral to the digestion and absorption of dietary peptides in the intestine.