A Glycosyltransferase from Sulfolobus solfataricus MT-4 Exhibits Poly(ADP-ribose) Glycohydrolase Activity

Author(s): Anna De Maio, Elena Porzio, Rosa D’Angelo, Sergio Rotondo, Anna Rita Bianchi, Elena Confalone, Raffaele Raucci, Emiliana Natale, Maria Rosaria Faraone-Mennella

Journal Name: Current Proteomics

Volume 12 , Issue 4 , 2015


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Abstract:

Anti-poly(ADP-ribose) glycohydrolase immunoblotting of a lysate from Sulfolobus solfataricus (strain MT-4) cells showed a main intense signal close to the 37 kDa protein marker. The immunoreactive protein was purified by electroelution and showed a hydrolysing activity towards oligomers (1- 6 residues) of ADP-ribose similar to eukaryotc poly(ADP-ribose) glycohydrolase. This protein was characterized as it regards enzymatic inhibition by adenosine diphosphate- (hydroxymethyl)pyrrolidine-3,4-diol, a known inhibitor of eukaryotic poly(ADP-ribose) glycohydrolase, and by analysis of reaction products. ADP-ribose polymer electrophoresis and thin layer chromatography clearly showed that the enzyme was able to monomerize Sulfolobus solfataricus MT-4 (ADP-ribose)1-6, an oligomer recognized also by eukaryotic poly (ADP-ribose) glycohydrolases.

Edman degradation of the purified protein allowed to determine a short N-terminal sequence: Met-Ile-Ser-Val-Ala. This pentapeptide was used for a blast search towards Sulfolobus solfataricus genomes. It gave evidence of a 40 kDa-protein present only in two strains (P2 and 98/2) of Sulfolobus solfataricus. Oligonucleotide primers drawn on the cDNA of human poly(ADP-ribose) glycohydrolase gave a fragment of the corresponding Sulfolobus solfataricus MT-4 gene overlapping the sequences from the genomes of Sulfolobus solfataricus P2 and 98/2. Translation of the sequence confirmed the occurrence of a region with some amino acids matching the human poly(ADP-ribose) glycohydrolase “signature”.

Keywords: Crenarchaeon, hyperthermophile, poly(ADP-ribose), poly(ADP-ribose) glycohydrolase, Sulfolobus; thermoprotein.

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Article Details

VOLUME: 12
ISSUE: 4
Year: 2015
Published on: 18 January, 2016
Page: [253 - 263]
Pages: 11
DOI: 10.2174/157016461204160119161543
Price: $25

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