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An orthogonal purification strategy for isolating crosslinked domains of modular synthases.

ABSTRACT: Chemo-enzymatic methods for covalently crosslinking carrier proteins with partner enzymes within modular synthases hold promise for elucidating and engineering metabolic pathways. Our efforts to crystallize the ACP-KS complexes of fatty acid synthases have been complicated by difficulties in the purification of the crosslinked complex from the other proteins in the reaction. Here we present a solution that employs an orthogonal purification strategy to achieve the quantity and level of purity necessary for further studies of this complex.

SUBMITTER: Haushalter RW

PROVIDER: S-EPMC3269958 | biostudies-literature | 2008 May

REPOSITORIES: biostudies-literature

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An orthogonal purification strategy for isolating crosslinked domains of modular synthases.

Haushalter Robert W RW Worthington Andrew S AS Hur Gene H GH Burkart Michael D MD

Bioorganic & medicinal chemistry letters 20080111 10

Chemo-enzymatic methods for covalently crosslinking carrier proteins with partner enzymes within modular synthases hold promise for elucidating and engineering metabolic pathways. Our efforts to crystallize the ACP-KS complexes of fatty acid synthases have been complicated by difficulties in the purification of the crosslinked complex from the other proteins in the reaction. Here we present a solution that employs an orthogonal purification strategy to achieve the quantity and level of purity ne ...[more]

PMID: 18249538

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