Date of Award


Degree Type


Degree Name

Doctor of Philosophy (PhD)



First Advisor

Kathryn B. Grant - Chair

Second Advisor

Alfons L. Baumstark

Third Advisor

Dabney W. Dixon


The development of new reagents to efficiently cleave peptides and proteins has become increasingly important for protein structural studies and other applications. However, this has proved to be a very challenging task due to the extreme stability of the peptide amide bond. Transition metal complexes cleave proteins and peptides through either oxidative or hydrolytic pathways. However, hydrolytic cleavage is preferred over oxidative cleavage, because the latter process produces irreversibly modified peptide fragments. Metal-assisted peptide hydrolysis is introduced in Chapter I. The metals Ce(IV), Co(II), Co(III), Cu(II), Fe(III), Mo(IV), Ni(II), Pd(II), Pt(II), Zn(II), and Zr(IV) are described as promising non-enzymatic hydrolysis reagents. In Chapter II, Zr(IV)-assisted hydrolysis of the dipeptide Gly-Gly and of its N- and C- blocked analogs is described. The highest levels of cleavage were observed at pH values ranging from 4.4 to 4.7. When the pH was raised to ~ 7.0, hydrolysis yields were decreased and amounts of zirconium precipitation were increased proportionately. Zirconium(IV)-assisted peptide hydrolysis in the presence of 4,13-diaza-18-crown-6 is reported in Chapter III. The goal of this work was to use an azacrown ether to reduce Zr(IV) precipitation and enhance levels of hydrolysis at neutral pH. An experiment in which 16 glycine containing dipeptides were hydrolyzed by Zr(IV) and by Zr(IV)/4,13-diaza-18-crown-6 indicated that 4,13-diaza-18-crown-6 markedly enhanced the reactivity of Zr(IV) under near physiological conditions. Because Zr(IV) precipitation was not reduced in these reactions, we proposed that hydrolysis of peptides by Zr(IV)/4,13-diaza-18-crown-6 might be heterogeneous in nature. In Chapter IV, seventeen macrocyclic and open-chain Zr(IV) ligands were compared in order to gain mechanistic insights that would enable hydrolysis yields at neutral pH to be further improved. While the macrocyclic ligands 4,13-diaza-18-crown-6 and 4,10-trioxa-7,13-diazacyclopentadecane tended to produce higher levels of Zr(IV)-assisted dipeptide cleavage, it was not necessary to have a ring structure to enhance Zr(IV) reactivity. With respect to the open-chain ligands, the potential ability to form multiple chelate rings appeared to coincide with reduced levels of Zr(IV) precipitation as well as with reduced levels of dipeptide hydrolysis. In Chapter V, a summary of our results and conclusions is presented.