Abstract: Methods for making and using substrates of deamidase of prokaryotic ubiquitin-like protein (Dop) are described herein. More particularly, modified prokaryotic ubiquitin-like protein (Pup) and functional fragments thereof that serve as exemplary Dop substrates are described and encompassed herein. Screening methods to identify modulators of Dop and Pup activity and use of modulators identified thereby are also described. Methods of using modulators that are identified as inhibitors of Dop and Pup activity for treating diseases/conditions associated with Mycobacterium tuberculosis (Mtb) infection, such as tuberculosis and leprosy, are also envisioned.

Abstract: Methods for making and using substrates of deamidase of prokaryotic ubiquitin-like protein (Dop) are described herein. More particularly, modified prokaryotic ubiquitin-like protein (Pup) and functional fragments thereof that serve as exemplary Dop substrates are described and encompassed herein. Screening methods to identify modulators of Dop and Pup activity and use of modulators identified thereby are also described. Methods of using modulators that are identified as inhibitors of Dop and Pup activity for treating diseases/conditions associated with Mycobacterium tuberculosis (Mtb) infection, such as tuberculosis and leprosy, are also envisioned.

Abstract: The present invention relates to prokaryotic ubiquitin-like protein (Pup) nucleic acid sequences and Pup amino acid sequences encoded therefrom. Also encompassed are antibodies that are immunologically specific for Pup. Methods directed to isolation and identification of pupylated substrates that utilize the conjugated Pup as an affinity tag are also included. Pupylated substrates that are identified using the method and reagents of the present invention provide tools useful for the identification of additional components of prokaryotic proteasomal machinery. Modulators of Pup activity and methods for identifying such modulators are also encompassed herein.

Abstract: The present invention relates to methods of treating Mycobacterium pathogen infection in a subject that involve: inhibiting proteasomal activity in a pathogen under conditions effective to make the pathogen susceptible to antibacterial host defenses; inhibiting enzyme activity in a pathogen under conditions effective to make the pathogen susceptible to antibacterial host defenses, where the enzyme is a DNA repair enzyme or a flavin-like co-factor synthesis enzyme, or inhibiting proteasomal and enzyme activity under conditions to make the pathogen susceptible to antibacterial host defenses. The present invention also relates to methods for screening compounds that inhibit proteasomal and protease activity, DNA repair enzyme activity, or flavin-like co-factor synthesis enzyme activity, where the inhibitory compounds have an ability to sensitize bacteria to the antibacterial effects of oxidative/nitrosative stress.

Abstract: The present invention relates to methods of treating pathogen infection in a subject that involve: inhibiting proteasomal activity in a pathogen under conditions effective to make the pathogen susceptible to antibacterial host defenses; inhibiting enzyme activity in a pathogen under conditions effective to make the pathogen susceptible to antibacterial host defenses, where the enzyme is a DNA repair enzyme or a flavin-like co-factor synthesis enzyme, or inhibiting proteasomal and enzyme activity under conditions to make the pathogen susceptible to antibacterial host defenses. The present invention also relates to methods for screening compounds that inhibit proteasomal and protease activity, DNA repair enzyme activity, or flavin-like co-factor synthesis enzyme activity, where the inhibitory compounds have an ability to sensitize bacteria to the antibacterial effects of oxidative/nitrosative stress.