RESUMO
Polyprenol phosphate phosphoglycosyl transferases (PGTs) catalyze the first membrane-committed step in assembly of essential glycoconjugates. Currently there is no structure-function information to describe how monotopic PGTs coordinate the reaction between membrane-embedded and soluble substrates. We describe the structure and mode of membrane association of PglC, a PGT from Campylobacter concisus. The structure reveals a unique architecture, provides mechanistic insight and identifies ligand-binding determinants for PglC and the monotopic PGT superfamily.
Assuntos
Campylobacter/enzimologia , Membrana Celular/enzimologia , Glicosiltransferases/química , Fosfatos/química , Catálise , Domínio Catalítico , Clonagem Molecular , Cisteína/química , Glicoconjugados/química , Ligantes , Mutação , Fosforilação , Domínios Proteicos , Dobramento de Proteína , Estrutura Secundária de Proteína , Relação Estrutura-Atividade , Especificidade por SubstratoRESUMO
For non-invasive imaging of the prostate cancer, we synthesized 7 alpha-fluoro-17 alpha-methyl 5 alpha-dihydrotestosterone ([(18)F]FMDHT) for androgen receptor mediated PET imaging. Preliminary in vitro and in vivo evaluations of this compound show promise. We designed and implemented a remote controlled system for reliable, efficient, and safe handling of radioactivity during the radiochemical synthesis of [(18)F]FMDHT. The key features of this report are the microwave assisted radiochemical synthesis, increased radiochemical yields, improved radiochemical purity, reduced overall synthesis time, and remote controlled automation of the entire synthesis. The overall synthesis using microwave reaction took 60-70 min and provided the desired product in 20-30% radiochemical yields with >99% radiochemical purity.