Structural and Functional Characterization of Staphylococcus aureus Enolase

Inhaltsverzeichnis (Table of Contents)

• Introduction
• Materials and Methods
  • Materials
  • Methods
• Results
  • Overexpression and Purification of Recombinant Enolase
  • Crystallization and Structure Determination of Staphylococcus aureus Enolase
  • In silico Analysis of Enolase Active Site and Structure
  • Kinetic and Thermodynamic Characterization of Enolase Activity
  • Inhibition Studies
  • Structural and Functional Characterization of Enolase Variants
• Discussion
• Conclusion

Zielsetzung und Themenschwerpunkte (Objectives and Key Themes)

This Ph.D. thesis aims to provide a comprehensive structural and functional characterization of Staphylococcus aureus enolase, an essential enzyme involved in glycolysis. The study investigates the enzyme's structure, active site, catalytic activity, and interactions with potential inhibitors. This research seeks to understand the molecular mechanisms underlying enolase function and identify potential targets for novel therapeutic strategies.

• Structural and Functional Characterization of Staphylococcus aureus Enolase
• Identification of Enolase Active Site and Interactions with Potential Inhibitors
• Analysis of Enolase Activity and Kinetic Parameters
• Investigation of Enolase Variants and Their Impact on Function
• Potential Applications in Drug Development and Therapeutic Strategies

Zusammenfassung der Kapitel (Chapter Summaries)

• Introduction: This chapter provides an overview of Staphylococcus aureus, a significant human pathogen, and its key metabolic pathways, particularly glycolysis. The role of enolase in glycolysis is highlighted, emphasizing its potential as a therapeutic target.
• Materials and Methods: This chapter details the materials and methods employed in the study, including recombinant protein expression, purification techniques, crystallization protocols, X-ray diffraction data collection and analysis, enzyme activity assays, kinetic studies, and inhibition experiments.
• Results: This chapter presents the results of the experimental investigations. It includes information on the overexpression and purification of recombinant enolase, the determination of its crystal structure, the in silico analysis of the active site and structural features, the kinetic and thermodynamic characterization of enolase activity, the identification and evaluation of potential inhibitors, and the structural and functional analysis of enolase variants.
• Discussion: This chapter interprets the results presented in the previous chapter, discussing the implications of the structural and functional insights for enolase activity and potential therapeutic applications.

Schlüsselwörter (Keywords)

This Ph.D. thesis primarily focuses on the structural and functional characterization of Staphylococcus aureus enolase, a key glycolytic enzyme. The research explores the enzyme's active site, kinetic properties, and interactions with potential inhibitors. Key themes include enzyme structure and function, protein crystallization, X-ray diffraction, catalytic activity, inhibition studies, and potential therapeutic applications. This work aims to contribute to the understanding of enolase as a potential drug target and to facilitate the development of novel therapeutic strategies against Staphylococcus aureus infections.