Kinetic characterization of bifunctional thymidylate synthase-dihydrofolate reductase (TS-DHFR) from Cryptosporidium hominis: a paradigm shift for ts activity and channeling behavior. (1/3)

This study presents a kinetic characterization of the recently crystallized bifunctional thymidylate synthasedihydrofolate reductase (TS-DHFR) enzyme from the apicomplexa parasite, Cryptosporidium hominis. Our study focuses on determination of the C. hominis TS-DHFR kinetic mechanism, substrate channeling behavior, and domain-domain communication. Unexpectedly, the unique mechanistic features of C. hominis TS-DHFR involve the highly conserved TS domain. At 45 s(-) (1), C. hominis TS activity is 10-40-fold faster than other TS enzymes studied and a new kinetic mechanism was required to simulate C. hominis TS behavior. A large accumulation of dihydrofolate produced at TS and a lag in product formation at DHFR were observed. These observations make C. hominis TS-DHFR the first bifunctional TS-DHFR enzyme studied for which there is clear evidence against dihydrofolate substrate channeling. Furthermore, whereas with Leishmania major TS-DHFR there are multiple lines of evidence for domain-domain communication (ligand binding at one active site affecting activity of the other enzyme), no such effects were observed with C. hominis TS-DHFR.  (+info)

Issue a boil-water advisory or wait for definitive information? A decision analysis. (2/3)

OBJECTIVE: Study the decision to issue a boil-water advisory in response to a spike in sales of diarrhea remedies or wait 72 hours for the results of definitive testing of water and people. METHODS: Decision analysis. RESULTS: In the base-case analysis, the optimal decision is test-and-wait. If the cost of issuing a boil-water advisory is less than 13.92 cents per person per day, the optimal decision is to issue the boil-water advisory immediately. CONCLUSIONS: Decisions based on surveillance data that are suggestive but not conclusive about the existence of a disease outbreak can be modeled.  (+info)

Back to basics: a revealing secondary reduction of the mitochondrial protein import pathway in diverse intracellular parasites. (3/3)

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