Purification and partial physical-chemical characterization of a new bovine trypsin proteoform (zeta-trypsin)

Fabiano Torres Cruz; Dayanne Pinho Rosa; Antonio Victor Baioco Vasconcelos; Jamil Silvano de Oliveira; Lucas Bleicher; Alexandre Martins Costa Santos

doi:10.1016/j.ijbiomac.2024.131860

Purification and partial physical-chemical characterization of a new bovine trypsin proteoform (zeta-trypsin)

Int J Biol Macromol. 2024 Apr 24;268(Pt 2):131860. doi: 10.1016/j.ijbiomac.2024.131860. Online ahead of print.

Authors

Fabiano Torres Cruz¹, Dayanne Pinho Rosa², Antonio Victor Baioco Vasconcelos¹, Jamil Silvano de Oliveira³, Lucas Bleicher⁴, Alexandre Martins Costa Santos⁵

Affiliations

¹ Pos-Graduate Program of Biotechnology - Federal University of Espírito Santo, Vitória, ES, Brazil.
² Pos-Graduate Program of Biochemistry - Federal University of Espírito Santo, Vitória, ES, Brazil.
³ Department of Biochemistry and Immunology - Federal University of Minas Gerais, Belo Horizonte, MG, Brazil.
⁴ Department of Biochemistry and Immunology - Federal University of Minas Gerais, Belo Horizonte, MG, Brazil; Pos-Graduate at Biochemistry and Immunology - Federal University of Minas Gerais, Belo Horizonte, MG, Brazil.
⁵ Pos-Graduate Program of Biotechnology - Federal University of Espírito Santo, Vitória, ES, Brazil; Pos-Graduate Program of Biochemistry - Federal University of Espírito Santo, Vitória, ES, Brazil. Electronic address: alexandre.santos@ufes.br.

PMID: 38670206
DOI: 10.1016/j.ijbiomac.2024.131860

Abstract

Recent advancements in enzyme research have unveiled a new proteoform of bovine trypsin, expanding our understanding of this well-characterized enzyme. While generally similar to other trypsins, this novel proteoform comprises three polypeptide chains, marking a significant difference in activity, kinetic properties, and conformational stability. Compared with the already known bovine trypsin proteoforms, the results showed a lower: activity, k_cat and k_cat.K_M^-1 and protein 'foldedness' ratio for the new proteoform. Molecular autolysis, a common feature in trypsin and chymotrypsin, has been explored through comparative physical chemistry properties with other proteoforms. This new proteoform of trypsin not only enriches the existing enzyme repertoire but also promises to shed light on the intricate physiological pathway for enzyme inactivation. Our results suggest that the new trypsin proteoform is one of the likely final pathways for enzyme inactivation in a physiological environment. This discovery opens up new avenues for further research into the functional implications of this new trypsin proteoform.

Keywords: Molecular autolysis; Pathway inactivation; Trypsin proteoform.