Bacterial Luciferase: Studies of Proteolytic Inactivation and Ligand Binding

The mechanism of proteolytic inactivation of luciferase was examined. Proteolytic inactivation in vitro was found to be concomittant with: (1) the loss of enzymatic activity; (2) altered enzyme affinity for substrate and product; (3) changes in enzyme secondary structure; and (4) the fragmentation of the (alpha) subunit into a discrete series of peptides, denoted the (gamma) and (delta) families. The region of the (alpha) subunit which was susceptible to proteolytic clipping was termed the Proteinase Labile Region and was observed to be a conserved structural feature in luciferases from distinct bacterial species. A preliminary study of the inactivation in vivo of the luciferase from Vibrio harveyi revealed the existence of isolatable antigenically cross-reactive peptide fragments bearing resemblance to peptides produced during proteolytic inactivation in vitro. The existence of fragments in vivo suggested that proteolysis in vitro may, in certain respects, parallel proteolytic events in vivo.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Holzman, Thomas Fredric

• Chemistry, Biochemistry

Identifier

(UMI)AAI8309959

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/70519