Date of this Version
Meyer, A.R. 2021. Structure of Clostridium perfringens Type IV Pili. Undergraduate Honors Thesis. University of Nebraska - Lincoln.
Type IV pili (T4P) are thin, hair-like bacterial appendages composed of protein subunits polymerized into a helical fiber. T4P perform diverse functions such as host cell adhesion, biofilm formation, natural competence, and twitching motility. While T4P are well characterized in Gram-negative bacteria, they have more recently been found in Gram-positive bacteria as well. In this work we aimed to solve the crystal structure of the type IV major pilin protein PilA2 from Clostridium perfringens, the predominant pilus subunit which makes up about 99% of the pilus fiber. We report expression, purification, and crystallization conditions which are sufficient for X-ray diffraction and data collection but lack sufficient resolution for structure elucidation at this time. Additionally, we display ab initio models of C. perfringens PilA2 that may inform future crystallization constructs and provide a template for molecular replacement phasing of C. perfringens PilA2 X-ray diffraction data. We believe successful structural determination will shed light on a poorly understood class of bacterial appendages, providing a model of T4P in similar Gram-positive species that may inform understanding of disease progression and therapeutic intervention in clostridial infections.