Analysis of catalytic properties of tripeptidyl peptidase I (TTP-I), a serine carboxyl lysosomal protease, and its detection in tissue extracts using selective FRET peptide substrate

Peptides
Marcia Y KondoMaria A Juliano

Abstract

Tripeptidyl peptidase I (TPP-I), also named ceroid lipofuscinosis 2 protease (CLN2p), is a serine carboxyl lysosomal protease involved in neurodegenerative diseases, and has both tripeptidyl amino- and endo- peptidase activities under different pH conditions. We developed fluorescence resonance energy transfer (FRET) peptides using tryptophan (W) as the fluorophore to study TPP-I hydrolytic properties based on previous detailed substrate specificity study (Tian Y. et al., J. Biol. Chem. 2006, 281:6559-72). Tripeptidyl amino peptidase activity is enhanced by the presence of amino acids in the prime side and the peptide NH2-RWFFIQ-EDDnp is so far the best substrate described for TPP-I. The hydrolytic parameters of this peptide and its analogues indicated that the S4 subsite of TPP-I is occluded and there is an electrostatic interaction of the positively charged substrate N-terminus amino group and a negative locus in the region of the enzyme active site. KCl activated TPP-I in contrast to the inhibition by Ca(2+) and NaCl. Solvent kinetic isotope effects (SKIEs) show the importance of the free N-terminus amino group of the substrates, whose absence results in a more complex solvent-dependent enzyme: substrate interaction and cata...Continue Reading

References

Dec 29, 1967·Biochemical and Biophysical Research Communications·N AbramowitzA Berger
Apr 20, 1967·Biochemical and Biophysical Research Communications·I Schechter, A Berger
Jul 11, 1998·Biochimica Et Biophysica Acta·D Vines, M J Warburton
Oct 23, 2002·Archives of Histology and Cytology·Masato KoikeYasuo Uchiyama
Dec 19, 2002·The Journal of Biological Chemistry·Adam A GolabekKrystyna E Wisniewski
Jan 2, 2004·The Journal of Biological Chemistry·Peter WujekAdam A Golabek
May 15, 2004·The Journal of Biological Chemistry·Adam A GolabekElizabeth Kida
Apr 16, 2008·The Journal of Biological Chemistry·Adam A GolabekElizabeth Kida
May 6, 2010·The Journal of Biological Chemistry·Márcia Y KondoIuri E Gouvea

❮ Previous
Next ❯

Citations


❮ Previous
Next ❯

Related Concepts

Related Feeds

Batten Disease

Batten Disease is a group of nervous system disorders known as neuronal ceroid lipofuscinosis. This feed focuses on neurobiological and neuropathological aspects of this disease.

Related Papers

European Journal of Paediatric Neurology : EJPN : Official Journal of the European Paediatric Neurology Society
Mohammed A JunaidRaju K Pullarkat
Archives of Biochemistry and Biophysics
Luciano PuzerAdriana K Carmona
© 2022 Meta ULC. All rights reserved