- Title Information
- Title
- Molecular characterization of allosteric signaling in D-dopachrome tautomerase
- Type of Resource (primo)
- dissertations
- Name:
Personal
- Name Part
- Chen, Emily
- Role
- Role Term:
Text
- creator
- Name:
Personal
- Name Part
- Jogl, Gerwald
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Fawzi, Nicolas
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Monaghan, Sean
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Smith, Colin
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Lisi, George
- Role
- Role Term:
Text
- Advisor
- Name:
Corporate
- Name Part
- Brown University. Department of Molecular Biology, Cell Biology and Biochemistry
- Role
- Role Term:
Text
- sponsor
- Origin Information
- Copyright Date
- 2022
- Physical Description
- Extent
- , None p.
- digitalOrigin
- born digital
- Note:
thesis
- Thesis (Ph. D.)--Brown University, 2022
- Genre (aat)
- theses
- Abstract
- Macrophage migration inhibitory factor (MIF) and its homolog MIF-2 are cytokine-like
proteins with critical roles in immunomodulation that are upregulated in various inflammatory
diseases. These proteins are broadly and constitutively expressed and bind CD74 to induce the
expression of pro-inflammatory cytokines, initiate ERK1/2 signaling, and to recruit leukocytes.
Commercial inhibitors of MIF target an enzymatic site, which has been demonstrated to alter
CD74-mediated activity when modified by mutation and inhibitor binding. However, their
therapeutic efficacy is hindered by a lack of molecular-level characterization of the effects of
inhibitor binding.
A dynamic relay connecting the MIF catalytic site to an allosteric site at Tyr99 was
previously reported. Its connectivity to the N- and C-terminus was determined by molecular
simulations and confirmed by experimentation where mutation at this site altered enzymatic
activity and CD74 activation. The structural conservation and functional overlap of MIF and MIF-2
prompted the question of whether this dynamic regulatory network is conserved in MIF-2.
Nuclear magnetic resonance (NMR) spectroscopy, tautomerase assays, and circular
dichroism was used in combination with X-ray crystallography, in vivo mouse models, and
molecular simulations from collaborators to present the structural, dynamic, and functional
consequences of mutating allosteric residues. Chapter Two details the N-terminal portion from
Pro1 to Phe100, the allosteric site in MIF-2. Chapter Three investigates the roles of Ser62 and
Phe100 in the network. Chapter Four defines the C-terminal segment continuing from Phe100 to
proposed CD74-activating residues based on homology to MIF. In each chapter, mutations of
residues in the allosteric network caused a range of structural and dynamic perturbations such that
flexibility correlated negatively with enzymatic activity. Further, mutations at the N-terminus
result in structural and dynamic perturbations at the C-terminus, and vice versa, demonstrating
allosteric reciprocity. This thesis establishes that dynamic communication is preserved in MIF-2
despite differences in primary sequence, defining a complete network from the N-terminal
enzymatic site to C-terminal CD74-activating residues that can be methodically targeted for
functional control of MIF family-induced inflammation.
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01040333")
- Topic
- Nuclear magnetic resonance spectroscopy
- Subject
- Topic
- Structural biology
- Subject
- Topic
- Allostery
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01742718")
- Topic
- Macrophage migration inhibitory factor
- Language
- Language Term (ISO639-2B)
- English
- Record Information
- Record Content Source (marcorg)
- RPB
- Record Creation Date
(encoding="iso8601")
- 20221018