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Characterization of Aplysia carboxypeptidase E
1Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
FEBS Letters
|May 19, 1997
Summary
Researchers expressed Aplysia californica carboxypeptidase E (CPE) in a baculovirus system. The study confirmed this cDNA encodes a CPE-like enzyme found in Aplysia tissues.
Area of Science:
- Biochemistry
- Molecular Biology
- Neuroscience
Background:
- Carboxypeptidase E (CPE) plays a crucial role in synthesizing peptide hormones and neurotransmitters.
- A cDNA encoding a potential Aplysia californica CPE was recently reported.
Purpose of the Study:
- To investigate whether the Aplysia californica cDNA encodes a functional CPE-like enzyme.
- To characterize the biochemical properties and tissue distribution of the Aplysia CPE.
Main Methods:
- Expression of Aplysia californica cDNA in the baculovirus system.
- Biochemical assays to determine optimal pH, inhibition by chelating agents and sulfhydryl reagents, and effects of divalent cations.
- Western blot analysis using specific antisera to detect Aplysia CPE in tissue.
- Affinity chromatography for enzyme purification.
Main Results:
- The expressed Aplysia enzyme exhibits optimal activity at pH 5.5-6.5.
- Enzyme activity is inhibited by chelating agents and p-chloromercuriphenyl sulfonate, similar to rat CPE.
- Western blots confirm Aplysia CPE presence in atrial glands and ovotestis.
- The enzyme was successfully purified using affinity chromatography.
Conclusions:
- The cloned Aplysia californica cDNA represents a functional CPE-like enzyme.
- This enzyme is expressed in key Aplysia tissues, suggesting a conserved role in neuropeptide processing.