Structural basis on the dityrosyl-diiron radical cluster and the functional differences of human ribonucleotide reductase small subunits hp53R2 and hRRM2

Bingsen Zhou, Leila Su, Yate Ching Yuan, Frank Un, Norby Wang, Madhukar Patel, Bixin Xi, Shuya Hu, Yun Yen

Research output: Contribution to journalArticle

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Abstract

Ribonucleotide reductase (RNR) is an enzyme for the de novo conversion of ribonucleotides to deoxyribonucleotides. The two human RNR small subunits hRRM2 and hp53R2 share 83% sequence homology but show distinct expression patterns and function. Structural analyses of the oxidized form of hRRM2 and hp53R2 indicate that both proteins contain a conserved Gln127-hp53R2/Gln165-hRRM2 close to the dinuclear iron center and the essential tyrosine residue Tyr124-hp53R2/Tyr162- hRRM2 forms hydrogen bonds with the tyrosine and iron ligands, implying a critical role for the glutamine residue in assembling the dityrosyl-diiron radical cofactor. The present work also showed that Tyr221 in hRRM2, which is replaced by Phe183 in hp53R2, forms a hydrogen bond with Tyr162 to extend the hydrogen bond network from Gln165-hRRM2. Mutagenesis and spectroscopic experiments suggested that the tyrosine-to-phenylalanine switch at Phe183-hp53R2/Tyr221-hRRM2 could lead to differences in radical generation or enzymatic activity for hp53R2 and hRRM2. This study correlates the distinct catalytic mechanisms of the small subunits hp53R2 and hRRM2 with a hydrogen-bonding network and provides novel directions for designing and developing subunitspecific therapeutic agents for human RNR enzymes.

Original languageEnglish
Pages (from-to)1669-1679
Number of pages11
JournalMolecular Cancer Therapeutics
Volume9
Issue number6
DOIs
Publication statusPublished - Jun 1 2010
Externally publishedYes

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Ribonucleotide Reductases
Tyrosine
Hydrogen
Iron
Deoxyribonucleotides
Ribonucleotides
Enzymes
Hydrogen Bonding
Sequence Homology
Glutamine
Phenylalanine
Mutagenesis
Ligands
Proteins
Therapeutics

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Structural basis on the dityrosyl-diiron radical cluster and the functional differences of human ribonucleotide reductase small subunits hp53R2 and hRRM2. / Zhou, Bingsen; Su, Leila; Yuan, Yate Ching; Un, Frank; Wang, Norby; Patel, Madhukar; Xi, Bixin; Hu, Shuya; Yen, Yun.

In: Molecular Cancer Therapeutics, Vol. 9, No. 6, 01.06.2010, p. 1669-1679.

Research output: Contribution to journalArticle

Zhou, Bingsen ; Su, Leila ; Yuan, Yate Ching ; Un, Frank ; Wang, Norby ; Patel, Madhukar ; Xi, Bixin ; Hu, Shuya ; Yen, Yun. / Structural basis on the dityrosyl-diiron radical cluster and the functional differences of human ribonucleotide reductase small subunits hp53R2 and hRRM2. In: Molecular Cancer Therapeutics. 2010 ; Vol. 9, No. 6. pp. 1669-1679.
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abstract = "Ribonucleotide reductase (RNR) is an enzyme for the de novo conversion of ribonucleotides to deoxyribonucleotides. The two human RNR small subunits hRRM2 and hp53R2 share 83{\%} sequence homology but show distinct expression patterns and function. Structural analyses of the oxidized form of hRRM2 and hp53R2 indicate that both proteins contain a conserved Gln127-hp53R2/Gln165-hRRM2 close to the dinuclear iron center and the essential tyrosine residue Tyr124-hp53R2/Tyr162- hRRM2 forms hydrogen bonds with the tyrosine and iron ligands, implying a critical role for the glutamine residue in assembling the dityrosyl-diiron radical cofactor. The present work also showed that Tyr221 in hRRM2, which is replaced by Phe183 in hp53R2, forms a hydrogen bond with Tyr162 to extend the hydrogen bond network from Gln165-hRRM2. Mutagenesis and spectroscopic experiments suggested that the tyrosine-to-phenylalanine switch at Phe183-hp53R2/Tyr221-hRRM2 could lead to differences in radical generation or enzymatic activity for hp53R2 and hRRM2. This study correlates the distinct catalytic mechanisms of the small subunits hp53R2 and hRRM2 with a hydrogen-bonding network and provides novel directions for designing and developing subunitspecific therapeutic agents for human RNR enzymes.",
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