Expression, purification and crystallization of human 3′-phosphoadenosine-5′-phosphosulfate synthetase 1

Authors: Harjes, Stefan; Scheidig, Axel; Bayer, Peter

Source: Acta Crystallographica Section D, Volume 60, Number 2, 1 February 2004 , pp. 350-352(3)

Publisher: Blackwell Publishing

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Abstract:

3′-Phosphoadenosine-5′-phosphosulfate (PAPS) is used to incorporate sulfate into biomolecules. The human PAPS synthetase 1 catalyzes two steps leading from adenosine triphosphate (ATP) and sulfate to PAPS. The ATP sulfurylase domain catalyzes the formation of the intermediate adenosine-5′-phosphosulfate (APS). The APS kinase domain then adds a phosphate group to the 3′-ribose and releases PAPS. In this article, the recombinant expression, purification and crystallization of the full-length protein is described. In Escherichia coli the protein is only partly soluble and copurifies with GroEL. The pure protein migrates as a dimer in gel-filtration chromatography. It is moderately active, forming 25 nmol PAPS per minute per milligram. Crystals grow to 100 × 100 × 300 µm and diffract to 1.75 Å.

Keywords: 3′-phosphoadenosine-5′-phosphosulfate synthetase 1

Document Type: Research article

DOI: 10.1107/S0907444903027628

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