Efficient identification of phosphatidylserine-binding proteins by ORF phage display

Nora B. Caberoy, Yixiong Zhou, Gabriela Alvarado, Xianqun Fan, Wei Li

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

To efficiently elucidate the biological roles of phosphatidylserine (PS), we developed open-reading-frame (ORF) phage display to identify PS-binding proteins. The procedure of phage panning was optimized with a phage clone expressing MFG-E8, a well-known PS-binding protein. Three rounds of phage panning with ORF phage display cDNA library resulted in ∼300-fold enrichment in PS-binding activity. A total of 17 PS-binding phage clones were identified. Unlike phage display with conventional cDNA libraries, all 17 PS-binding clones were ORFs encoding 13 real proteins. Sequence analysis revealed that all identified PS-specific phage clones had dimeric basic amino acid residues. GST fusion proteins were expressed for 3 PS-binding proteins and verified for their binding activity to PS liposomes, but not phosphatidylcholine liposomes. These results elucidated previously unknown PS-binding proteins and demonstrated that ORF phage display is a versatile technology capable of efficiently identifying binding proteins for non-protein molecules like PS.

Original languageEnglish
Pages (from-to)197-201
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume386
Issue number1
DOIs
StatePublished - Aug 14 2009

Fingerprint

Bacteriophages
Phosphatidylserines
Open Reading Frames
Carrier Proteins
Display devices
Clone Cells
Gene Library
Liposomes
Basic Amino Acids
Phosphatidylcholines
Sequence Analysis
Proteins
Fusion reactions
Technology

Keywords

  • ORF phage display
  • Phage display
  • Phosphatidylcholine
  • Phosphatidylserine
  • Phosphatidylserine-binding protein

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Efficient identification of phosphatidylserine-binding proteins by ORF phage display. / Caberoy, Nora B.; Zhou, Yixiong; Alvarado, Gabriela; Fan, Xianqun; Li, Wei.

In: Biochemical and Biophysical Research Communications, Vol. 386, No. 1, 14.08.2009, p. 197-201.

Research output: Contribution to journalArticle

Caberoy, Nora B. ; Zhou, Yixiong ; Alvarado, Gabriela ; Fan, Xianqun ; Li, Wei. / Efficient identification of phosphatidylserine-binding proteins by ORF phage display. In: Biochemical and Biophysical Research Communications. 2009 ; Vol. 386, No. 1. pp. 197-201.
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