Investigation of Adenylate Kinases Isolated From Mesophilic and Thermophilic Members of the Archaeal Genus Methanococcus

Construction of chimeric proteins between M. voltae and M. jannaschii AKs along with site specific mutational analysis confirm the importance of specific stabilizing interactions identified by sequence comparisons, and allow the physical characteristic of these proteins to be significantly altered. Simple protein engineering increased the temperature optima of the Methanococcus voltae AK from 37$\sp\circ$C to 62$\sp\circ$C and its melting point from 68$\sp\circ$C to 89$\sp\circ$C while maintaining specific activity levels and approximately 90% of its original sequence. Analysis of the chimerical proteins also point strongly to the cooperative and non-additive nature of thermal stabilizing interactions within the methanococcal AKs. These studies also indicate that the molecular mechanisms which determine enzymatic temperature optima and overall protein stability are not directly related.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Haney, Paul Jeffrey

Contributors dc:contributor

• Konisky, Jordan

• Chemistry, Biochemistry

Language dc:language

eng

Identifier

(MiAaPQ)AAI9812611

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/86722