Effects of monovalent and divalent ions in factor xiii activation and crosslinking.

Author

Richard Laporca Lumata, University of LouisvilleFollow

Date on Master's Thesis/Doctoral Dissertation

8-2022

Document Type

Master's Thesis

Degree Name

M.S.

Department

Chemistry

Degree Program

Chemistry, MS

Committee Chair

Maurer, Muriel

Committee Member

Li, Ying

Committee Member

Wilson, Andrew

Committee Member

Menze, Michael

Author's Keywords

FXIII; monodansylcadaverine; transglutaminase; thrombin; Cab3

Abstract

Factor XIII (FXIII) is an emerging target for treating blood clotting and cardiovascular related diseases. FXIII can be activated non-proteolytically by the presence of elevated Ca²⁺ levels (2-100 mM) or proteolytically by thrombin-cleavage of the Activation Peptide along with low mM Ca²⁺. The studies herein utilized fluorescence to examine how monovalent and divalent ions influence the transglutaminase activity and conformation of FXIII. Monodansylcadaverine assays revealed that increasing ionic radius (Cs⁺ > K⁺ > Na⁺ > Li⁺) and increasing ionic strength (XCl^- levels and SO₄^2- > Cl^-) elevated FXIII-A transglutaminase activity. Intrinsic fluorescence studies revealed that only cations influenced FXIII conformation. For divalent salts, transglutaminase activity for MgSO₄ is ~3-fold higher than MgCl₂. Unlike transglutaminase 2, FXIII containing a mutation in the Cab3 calcium binding site (G262V) could not exhibit an improvement in activity. A FXIII Cab3 helix is proposed to be hindered which impacts non-proteolytically activated FXIII more than proteolytically activated FXIII. Both cations and anions influence FXIII’s ability to interact with substrates.

Recommended Citation

Lumata, Richard Laporca, "Effects of monovalent and divalent ions in factor xiii activation and crosslinking." (2022). Electronic Theses and Dissertations. Paper 3969.
https://doi.org/10.18297/etd/3969