Crystal structure of Saccharomyces cerevisiae 3′-phosphoadenosine- 5′-phosphosulfate reductase complexed with adenosine 3′,5′- bisphosphate

Zhihao Yu, Donna Lemongello, Irwin H. Segel, Andrew J Fisher

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Most assimilatory bacteria, fungi, and plants species reduce sulfate (in the activated form of APS or PAPS) to produce reduced sulfur. In yeast, PAPS reductase reduces PAPS to sulfite and PAP. Despite the difference in substrate specificity and catalytic cofactor, PAPS reductase is homologous to APS reductase in both sequence and structure, and they are suggested to share the same catalytic mechanism. Metazoans do not possess the sulfate reduction pathway, which makes APS/PAPS reductases potential drug targets for human pathogens. Here, we present the 2.05 Å resolution crystal structure of the yeast PAPS reductase binary complex with product PAP bound. The N-terminal region mediates dimeric interactions resulting in a unique homodimer assembly not seen in previous APS/PAPS reductase structures. The "pyrophosphate- binding" sequence 47TTAFGLTG54 defines the substrate 3′-phosphate binding pocket. In yeast, Gly54 replaces a conserved aspartate found in APS reductases vacating space and charge to accommodate the 3′-phosphate of PAPS, thus regulating substrate specificity. Also, for the first time, the complete C-terminal catalytic motif 244ECGIH 248 is revealed in the active site. The catalytic residue Cys245 is ideally positioned for an in-line attack on the β-sulfate of PAPS. In addition, the side chain of His248 is only 4.2 Å from the Sγ of Cys245 and may serve as a catalytic base to deprotonate the active site cysteine. A hydrophobic sequence 252RFAQFL257 at the end of the C-terminus may provide anchoring interactions preventing the tail from swinging away from the active site as seen in other APS/PAPS reductases.

Original languageEnglish (US)
Pages (from-to)12777-12786
Number of pages10
JournalBiochemistry
Volume47
Issue number48
DOIs
StatePublished - Dec 2 2008

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Phosphoadenosine Phosphosulfate
Yeast
Adenosine
Saccharomyces cerevisiae
Oxidoreductases
Crystal structure
Sulfates
Catalytic Domain
Yeasts
Substrate Specificity
Substrates
Phosphates
Sulfites
Pathogens
Fungi
Sulfur
Aspartic Acid
Cysteine
3'-phosphoadenylyl-5'-phosphosulfate reductase
Tail

ASJC Scopus subject areas

  • Biochemistry

Cite this

Crystal structure of Saccharomyces cerevisiae 3′-phosphoadenosine- 5′-phosphosulfate reductase complexed with adenosine 3′,5′- bisphosphate. / Yu, Zhihao; Lemongello, Donna; Segel, Irwin H.; Fisher, Andrew J.

In: Biochemistry, Vol. 47, No. 48, 02.12.2008, p. 12777-12786.

Research output: Contribution to journalArticle

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