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Cloning, expression and functional characterization of rat napsin.
V Schauer-Vukasinovic1, M B Wright, V Breu
1F. Hoffmann-La Roche Ltd., Pharma Division, Preclinical Research, Grenzacherstrasse 124, CH-4070 Basel, Switzerland.
Biochimica Et Biophysica Acta
|June 20, 2000
Summary
Researchers identified rat napsin, a protein highly expressed in the kidney, lung, and spleen. This protein demonstrated enzymatic activity when expressed in human embryonic kidney cells.
Area of Science:
- Molecular Biology
- Biochemistry
- Genomics
Background:
- Napsin, a proteinase, plays a role in biological processes.
- Understanding the expression and function of napsin is crucial.
Purpose of the Study:
- To identify and characterize the full-length cDNA clone for rat napsin.
- To investigate the expression pattern and enzymatic activity of rat napsin.
Main Methods:
- Homology search of the ZooSeq rat EST database.
- Northern blot analysis for mRNA expression.
- Western blot analysis for protein expression.
- Enzymatic activity assay in transfected HEK293 cells.
Main Results:
- A full-length cDNA clone coding for rat napsin was identified.
- Rat napsin mRNA transcripts were highly expressed in kidney, lung, and spleen.
- Rat napsin was confirmed as a 50-kDa, highly glycosylated, monomeric protein in kidney.
- Transfected HEK293 cells exhibited increased napsin enzymatic activity, inhibited by pepstatin.
Conclusions:
- Rat napsin is a highly expressed protein in specific rat organs.
- The identified napsin clone encodes a functional enzyme with pepstatin-sensitive activity.