Identification, expression, and purification of a unique stable domain from human HSPC144 protein

Ai Xin Song, Yong Gang Chang, Yong Guang Gao, Xiao Jing Lin, Yan Hong Shi, Dong Hai Lin, Qiu Hua Hang, Hong Yu Hu

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

HSPC144 is a newly identified gene in human CD34+ hematopoietic stem/progenitor cells. In this work, we have expressed and purified the 225-residue protein from Escherichia coli BL21 (DE3) and identified a stable fragment HSPC144-P (residues 44-225) by limited proteolysis method. The HSPC144-P fragment exhibits high stability with a little increase of secondary structure percentage as compared with the full-length protein. We anticipated that the N-terminally truncated protein possesses a more compact structure. By sequence analysis, the proteolytic fragment shares a great similarity with DUF589 domain, a previously identified domain with unknown function. This novel domain is highly conserved in Thy28 proteins and is worthy of structural and functional studies. We have subcloned this homologous domain from HSPC144 protein and purified to homogeneity for structure analysis. The 15N and 15N/13C-labeled DUF589 domain samples have been prepared successfully and determination of the NMR structure is in progress.

Original languageEnglish (US)
Pages (from-to)146-152
Number of pages7
JournalProtein Expression and Purification
Volume42
Issue number1
DOIs
StatePublished - Jul 2005

Bibliographical note

Funding Information:
This work was supported by grants from the 863 Hi-Tech program (2002BA711A13), the Chinese Academy of Sciences (KSCX1-SW-17) and the Shanghai Commission of Science and Technology (03JC14081).

Keywords

  • DUF589 domain
  • Expression
  • HSPC protein
  • Limited proteolysis
  • NMR analysis
  • Purification

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