Employing solution NMR for structural interrogation of the Btk SH2 domain to understand its role in kinase activation
Date
2023-12
Authors
Lowe, Jacques
Major Professor
Advisor
Andreotti, Amy
Sashital, Dipali
Underbakke, Eric
Nelson, Scott
Venditti, Vincenzo
Committee Member
Journal Title
Journal ISSN
Volume Title
Publisher
Altmetrics
Abstract
There are over 500 eukaryotic protein kinases encoded in the human genome, making up nearly 2% of the entire genome. Establishing their dominance, kinases regulate most of the biological processes that take place in the cell. Therefore, strict regulation of kinase activity is necessary for all cells to function normally and maintain a healthy state.
Kinases are often described as molecular switches that can interconvert between active and inactive states in response to an external stimulus. The switch can be as simple as turning the activity “on/off” like a light switch, or it can be more gradual and fine-tuned like a thermostat dial. The Tec family non-receptor tyrosine kinases are more like the gradually, fine-tuned thermostat, acting as key nodes within the signaling network, downstream of the antigen receptor. Tec family kinases work to transduce signals that lead to calcium mobilization, gene expression and cell proliferation.
Numerous studies have pursued the autoinhibited state of the kinases within non-receptor tyrosine kinase subfamilies, and so far, the field has developed a clear picture of the features of the inactive state- or “off” switch. More recently, efforts have also been made to describe the active state, however, there remains a gap in our understanding of the active state conformations of kinases. Factors known to mediate activation include phosphorylation, binding of allosteric regulators, and changes in cellular localization, yet the molecular basis of activation is still unknown for many kinases.
This dissertation focuses on the Tec kinase, Bruton’s tyrosine kinase (BTK), with a focus on positive regulation of this enzyme. Described herein are the various techniques that were used to gain insight into the structure and dynamics of BTK, with emphasis on an active fragment of the BTK kinase. Together, this work provides new structural information regarding the BTK active state, as well as highlighting the potential for allosteric control of BTK function and the development of novel, targeted therapies for treatment of varying immune disorders.
Series Number
Journal Issue
Is Version Of
Versions
Series
Academic or Administrative Unit
Biochem, Biophysics, and Molecular Biology
Type
article