Team:Washington/Gram Positive

Anthrax

Anthrax is a lethal disease caused by the bacteria Bacillus anthracis which can spread by ingestion, inhalation, or cutaneous contact with spores. Spores of B. anthracis are extremely resilient, remaining potent even in the harshest climates for multiple decades or centuries. This disease is not contagious, cannot be transferred from an infected organism, though bacterial spores can be transported in a multitude of ways and will infect potential victims.

CapD

Anthrax creates a poly-γ-D-glutamate (PDGA) capsule which prevents the immune system from recognizing it as a pathogen and performing phagocytosis to destroy the threat. Naturally, anthrax produces CapD which cleaves and anchors the capsule to its peptidoglycan, the outer coat of the cell membrane; however, research shows that an overexpression of CapD will cleave the capsule, removing the bacteria’s immunity to phagocytosis.

Goal

CapD is naturally a transpeptidase, favoring reactions with amino acids to perform its task, cleaving PDGA. However, CapD would be invaluable as a hydrolase, reacting with water to cleave PDGA, because of the high levels of water in the human body. If a mutant CapD could be engineered as an extremely efficient hydrolase, it is theorized that a single dose of concentrated CapD into the blood stream would easily decimate anthrax and nullifying its lethal properties.

Other

here's an outline from the 8/25 meeting. the pages are set up based on it so far but feel free to change them

• Background (this page)
  • Anthrax is bad
  • Your immune system can't destroy it while its coating is intact
  • Our protein takes off the coating
• Introduction/methods
  • transpeptidase --> hydrolase
  • protein engineering in foldit
  • protein purification
  • kunkel mutagenesis
  • michaelis-menton (sp?) is replacing mutant data?
  • protein expression vectors
• experimental results
  • relative activities of variants
  • do this page last because it will change as we go

something should point to Tom's vectors project... not sure where that goes best.

update 8/30:

• spend less time on kunkle mutagenesis
• divide process wheel, overall presentation, this wiki to "design --> build --> test"
• add youtube videos
• organize into treatment/therapeutics —> positive, negative