This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The proteasome is the major cellular protease. It is involved in the controlled degradation of proteins that regulate a wide variety of cellular processes, such as transcription, apoptosis, cell division and DNA repair. With an important role in homeostasis of so many proteins it is not surprising that observed increased proteasome activity (e.g. in multiple myelomas) or decreased proteasome activity (e.g. in many neurodegenerative diseases) is a pathological factor in many diseases. One determinant of cellular proteasome activity is the level of proteasomes in the cell. Thus, it is important to understand how cellular proteasome levels are regulated and how proteasome assembly is regulated. The long-term goal of this project is to understand the mechanisms of proteasome assembly. Recent work has identified four chaperones that facilitate the formation of proteasome regulatory particle (RP). Each chaperone binds to the C-domain of a specific AAA-ATPases located in the RP. Despite this similarity in function and binding properties, there is no sequence or structural conservation among these chaperones. The objective of the research proposed here is to understand the role each RP chaperone plays in the formation of proteasomes. We hypothesize that the chaperones act as templates for specific base subunits in the assembly of RP. Secondly, they regulate the order in which precursor complexes assemble in a spatial and temporal manner. We also hypothesize that the structural difference among the chaperones are important to accommodate as well as prevent a unique set of interactions for specific ATPases during the assembly process. We will use in vitro binding and reconstitution assays to study this. We furthermore will obtain structural information of the chaperones in combination with the C-domain they bind to. We expect that information from the experiments proposed here will provide a detailed understanding of the mechanisms the chaperones employ to assist proteasome assembly. This could potentially provide new drug targets and ultimately enable us to manipulate proteasome levels or activity for therapeutic means.

National Institute of Health (NIH)
National Center for Research Resources (NCRR)
Exploratory Grants (P20)
Project #
Application #
Study Section
National Center for Research Resources Initial Review Group (RIRG)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Kansas Lawrence
Schools of Pharmacy
United States
Zip Code
Pang, Xiao-Yan; Wang, Suya; Jurczak, Michael J et al. (2017) Retinol saturase modulates lipid metabolism and the production of reactive oxygen species. Arch Biochem Biophys 633:93-102
Jeanne Dit Fouque, Kevin; Garabedian, Alyssa; Porter, Jacob et al. (2017) Fast and Effective Ion Mobility-Mass Spectrometry Separation of d-Amino-Acid-Containing Peptides. Anal Chem 89:11787-11794
Alaofi, Ahmed; Farokhi, Elinaz; Prasasty, Vivitri D et al. (2017) Probing the interaction between cHAVc3 peptide and the EC1 domain of E-cadherin using NMR and molecular dynamics simulations. J Biomol Struct Dyn 35:92-104
McNiff, M L; Haynes, E P; Dixit, N et al. (2016) Thioredoxin fusion construct enables high-yield production of soluble, active matrix metalloproteinase-8 (MMP-8) in Escherichia coli. Protein Expr Purif 122:64-71
Johnson, Troy A; Mcleod, Matthew J; Holyoak, Todd (2016) Utilization of Substrate Intrinsic Binding Energy for Conformational Change and Catalytic Function in Phosphoenolpyruvate Carboxykinase. Biochemistry 55:575-87
Tucker, Jenifer K; McNiff, Michaela L; Ulapane, Sasanka B et al. (2016) Mechanistic investigations of matrix metalloproteinase-8 inhibition by metal abstraction peptide. Biointerphases 11:021006
Yadav, Rahul; Vattepu, Ravi; Beck, Moriah R (2016) Phosphoinositide Binding Inhibits Actin Crosslinking and Polymerization by Palladin. J Mol Biol 428:4031-4047
Gurung, Ritu; Yadav, Rahul; Brungardt, Joseph G et al. (2016) Actin polymerization is stimulated by actin cross-linking protein palladin. Biochem J 473:383-96
Budiardjo, S Jimmy; Licknack, Timothy J; Cory, Michael B et al. (2016) Full and Partial Agonism of a Designed Enzyme Switch. ACS Synth Biol 5:1475-1484
Gowthaman, Ragul; Miller, Sven A; Rogers, Steven et al. (2016) DARC: Mapping Surface Topography by Ray-Casting for Effective Virtual Screening at Protein Interaction Sites. J Med Chem 59:4152-70

Showing the most recent 10 out of 253 publications