Analysis of Bacterial ATP-Binding Cassette Transporters

Our experts have a keen interest in the study of liquid-liquid and liquid-solid phase transitions in bacterial cells. We have cutting-edge super-resolution imaging or infinite diffraction microscopy apparently combined with single molecule trafficking methods, and computer analysis platforms to analyze key microbial biomolecular condensates undergoing LLPS, as well as the formation and organization of biomolecular condensates within the intracellular space. Here, CD BioSciences is committed to analyzing intrinsically disordered bacterial ATP-binding cassette (ABC) transporters.

Introduction of Bacterial ATP-Binding Cassette Transporters

The ATP-binding cassette (ABC) transporter superfamily is present in all life from eukaryotes to prokaryotes and facilitates key biological processes across cell membranes by transferring various substrates from ions to proteins during ATP coupling. Mycobacterium tuberculosis ABC transporter Rv1747 belongs to a large superfamily of multisubunit permeases that transport a variety of molecules across biological membranes. It is a putative exporter of intermediates for cell wall biosynthesis and is important for the growth of Mycobacterium tuberculosis in infected hosts. This integral membrane protein (monomer) consists of two forkhead-associated (FHA) structural domains that are separated from each other by an essentially disordered junction, a cytoplasmic nucleotide-binding structural domain, and a transmembrane structural domain. Studies have shown that the FHA structural domain has the intrinsic ability to form spherical liquid-like condensates.

Fig. 1. Schematic representation of ATP-binding cassette (ABC) transporters. (Akhtar A A, et al., 2022)

Customized Services

The FHA domain of Rv1747 is a signaling protein module that mediates protein-protein interactions and regulates a variety of biological processes by binding to conserved phosphorylated threonine-containing (pT) interacting oligopeptides. Our technical team is dedicated to analyzing the phase separation and aggregation of Mycobacterium tuberculosis ABC transporter Rv1747.

CD BioSciences provides professional services to analyze the structural dynamics and biochemical functions of the intrinsically disordered ABC transporter Rv1747 in bacteria. Our experts have developed one-stop working models of STPKs/PstP allowing rapid and reversible activation/inactivation of Rv1747 upon initial exposure of infected macrophages to challenging environments.

• Characterization of ABC Transporter Rv1747 LLPS in Vitro
  Our laboratory has NMR to characterize the FHA-1 or FHA-2 structural domains. We provide turbidimetric, microscopic, and fluorescence spectroscopy platforms to analyze the interactions mediated by the regulatory module FHA structural domain of Rv1747 and the linker phosphorylated receptor threonine to characterize the intrinsic ability of Rv1747 to form condensates at high concentrations in vitro. We are also able to analyze the ability of non-phosphorylated Rv1747 to undergo phase separation.
  Our single-molecule localization microscopy (SMLM) platform is widely used to analyze higher-order nanoclusters of endogenous Mycobacterium tuberculosis Rv1747.
  
• Development of ABC Transporter Therapeutics
  ABC transporters have been implicated in the virulence and pathogenesis of a variety of pathogenic bacteria, such as input of essential nutrients, secretion of proteins and output of toxic xenobiotics. Our experts are committed to developing computational methods and high-throughput data mining approaches for the identification and characterization of unknown bacterial ABC transporters.
  In addition, we are developing novel ABC inhibitors and substrate mimics and exploring the potential of gene therapy to reverse ABC transporter protein-mediated multidrug resistance.

CD BioSciences offers a simple modular biomolecular platform to characterize the LLPS of intrinsically disordered ABC transporter Rv1747. We aim to analyze the molecular interactions that drive ABC transporter Rv1747 assembly in bacteria and develop ABC transporter therapeutics. If you have any special requirements for our services, please feel free to contact us.