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 Co-Chair (if applicable)

Li, Ying

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 SO42- > 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.

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