Site specific and reversible protein immobilization facilitated by a DNA binding fusion tag

Meng Zhong, Jun Fang, Yinan Wei

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


Due to the complexity and diversity of protein structures, site-specific protein immobilization has always been challenging. On the contrary, DNA immobilization is straightforward with well established chemical methods. Single-strand DNA binding protein (SSB) binds tightly to single-stranded DNA (ssDNA). Herein, we investigated the feasibility of using SSB as a fusion tag to facilitate site-specific and reversible immobilization of target proteins. As a model system, we constructed a fusion protein by joining a superfolder green fluorescent protein (sfGFP) with SSB. The fluorescence emission and ssDNA binding affinity of the fusion protein were compared separately with those of the individual modules. Both modules fully retained their properties in the fusion construct. Next, we covalently attached ssDNA (dT37) to a supporting matrix through either amine or thiol functionalization. The attached ssDNA mediated reversible sfGFP-SSB immobilization. The immobilized protein could be released through changes of conditions including pH, concentration of divalent cations, and the presence of the complementary dA35 oligonucleotide.

Original languageEnglish
Pages (from-to)1177-1182
Number of pages6
JournalBioconjugate Chemistry
Issue number7
StatePublished - Jul 21 2010

Bibliographical note

Funding Information:
The authors would like to thank the Ministerio de Econom?a y Competitividad (Project No. PCIN-2013-183), the Sociedad Espa?ola de Cat?lisis (SECAT Introducci?n a la Investigaci?n en Cat?lisis grant) and the Spanish government (Grant No. FPU13/00727) for their financial support.

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry


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