Dimerization of human XPA and formation of XPA2-RPA protein complex

Zheng guan Yang, Yang Liu, Leslie Y. Mao, Jian-Ting Zhang, Yue Zou

Research output: Contribution to journalArticle

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Abstract

XPA plays an important role in the DNA damage recognition during human nucleotide excision repair. Here we report that the XPA is a homodimer either in the free state or as a complex with human RPA in solution under normal conditions. The human XPA protein purified from baculovirus-infected sf21 insect cells has a molecular mass of 36 317 Da, as determined by mass spectroscopy. However, the apparent molecular mass of XPA determined by the native gel filtration chromatography was about 71 kDa, suggesting that XPA is a dimer. This observation was supported by a native PFO-PAGE and fluorescence spectroscopy analysis. XPA formed a dimer (XPA2) in a broad range of XPA and NaCl concentrations, and the dimerization was not due to the disulfide bond formation. Furthermore, a titration analysis of the binding of XPA to the human RPA indicated that it was the XPA2 that formed the complex with RPA. Finally, the difference between the mass spectrometric and the calculated masses of XPA implies that the protein contains posttranslational modifications. Taken together, our data suggest that the dimerization of XPA may play an important role in the DNA damage recognition of nucleotide excision repair.

Original languageEnglish
Pages (from-to)13012-13020
Number of pages9
JournalBiochemistry
Volume41
Issue number43
DOIs
StatePublished - Oct 29 2002

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Dimerization
Molecular mass
Dimers
Repair
Nucleotides
DNA Repair
DNA Damage
DNA
Fluorescence spectroscopy
Chromatography
Titration
Disulfides
Native Polyacrylamide Gel Electrophoresis
Sf9 Cells
Proteins
Baculoviridae
Gels
Fluorescence Spectrometry
Spectroscopy
Post Translational Protein Processing

ASJC Scopus subject areas

  • Biochemistry

Cite this

Yang, Z. G., Liu, Y., Mao, L. Y., Zhang, J-T., & Zou, Y. (2002). Dimerization of human XPA and formation of XPA2-RPA protein complex. Biochemistry, 41(43), 13012-13020. https://doi.org/10.1021/bi026064z

Dimerization of human XPA and formation of XPA2-RPA protein complex. / Yang, Zheng guan; Liu, Yang; Mao, Leslie Y.; Zhang, Jian-Ting; Zou, Yue.

In: Biochemistry, Vol. 41, No. 43, 29.10.2002, p. 13012-13020.

Research output: Contribution to journalArticle

Yang, ZG, Liu, Y, Mao, LY, Zhang, J-T & Zou, Y 2002, 'Dimerization of human XPA and formation of XPA2-RPA protein complex', Biochemistry, vol. 41, no. 43, pp. 13012-13020. https://doi.org/10.1021/bi026064z
Yang ZG, Liu Y, Mao LY, Zhang J-T, Zou Y. Dimerization of human XPA and formation of XPA2-RPA protein complex. Biochemistry. 2002 Oct 29;41(43):13012-13020. https://doi.org/10.1021/bi026064z
Yang, Zheng guan ; Liu, Yang ; Mao, Leslie Y. ; Zhang, Jian-Ting ; Zou, Yue. / Dimerization of human XPA and formation of XPA2-RPA protein complex. In: Biochemistry. 2002 ; Vol. 41, No. 43. pp. 13012-13020.
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