Activation and processing of non-anchored yapsin 1 (Yap3p).

The Journal of Biological Chemistry
Niamh X CawleyY P Loh

Abstract

A C-terminally truncated form of yapsin 1 (yeast aspartic protease 3), the first member of the novel sub-class of aspartic proteases with specificity for basic residues (designated the Yapsins), was overexpressed and purified to apparent homogeneity, yielding approximately 1 microg of yapsin 1/g of wet yeast. N-terminal amino acid analysis of the purified protein confirmed that the propeptide was absent and that the mature enzyme began at Ala68. The mature enzyme was shown to be composed of approximately equimolar amounts of two subunits, designated alpha and beta, that were associated to each other by a disulfide bond. C-terminally truncated proyapsin 1 was also expressed in the baculovirus/Sf9 insect cell expression system and secreted as a zymogen that could be activated upon incubation at an acidic pH with an optimum at approximately 4.0. When expressed without its pro-region, it was localized intracellularly and lacked activity, indicating that the pro-region was required for the correct folding of the enzyme. The activation of proyapsin 1 in vitro exhibited linear kinetics and generated an intermediate form of yapsin 1 or pseudo-yapsin 1.

References

Mar 1, 1979·European Journal of Biochemistry·V B PedersenB Foltmann
Mar 25, 1992·Nucleic Acids Research·D GietzR H Schiestl
Jan 1, 1991·Enzyme·Y BourbonnaisD Y Thomas
Mar 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·R S FullerJ Thorner
Jan 1, 1988·Annual Review of Physiology·R S FullerJ Thorner
Jan 1, 1987·Journal of Cellular Biochemistry·J Tang, R N Wong
Nov 7, 1995·Proceedings of the National Academy of Sciences of the United States of America·H Komano, R S Fuller
Dec 1, 1994·Kidney International·T L ReudelhuberN G Seidah
Apr 14, 1995·The Journal of Biological Chemistry·H B van den HazelJ R Winther
Mar 10, 1995·The Journal of Biological Chemistry·O SeksekA S Verkman

❮ Previous
Next ❯

Citations

May 5, 2005·Applied and Environmental Microbiology·Marc W T Werten, Frits A de Wolf
Jun 10, 2011·Biotechnology and Bioengineering·Catarina I F SilvaMarc W T Werten
Oct 18, 2006·FEMS Yeast Research·Isabelle Gagnon-ArsenaultYves Bourbonnais
Dec 2, 1999·FEBS Letters·V Schauer-VukasinovicT Giller
Oct 1, 2010·Biomolecular Concepts·Ilya V DemidyukSergey V Kostrov
Feb 13, 2014·The Journal of Biological Chemistry·Martin LoiblSabine Strahl
Mar 4, 2000·Archives of Biochemistry and Biophysics·V Olsen, Y P Loh
Dec 12, 2002·Chemical Reviews·Philip N Bryan
Jan 17, 2007·Biochemistry·Kathryn E FisherPhilip N Bryan

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.