a screening of a library of t7 phage-displayed peptide identifies e2f-4 as an etoposide-binding protein库的筛选t7 phage-displayed肽识别e2f-4 etoposide-binding蛋白质.pdfVIP

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a screening of a library of t7 phage-displayed peptide identifies e2f-4 as an etoposide-binding protein库的筛选t7 phage-displayed肽识别e2f-4 etoposide-binding蛋白质.pdf

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a screening of a library of t7 phage-displayed peptide identifies e2f-4 as an etoposide-binding protein库的筛选t7 phage-displayed肽识别e2f-4 etoposide-binding蛋白质

Molecules 2011, 16, 4278-4294; doi:10.3390/molecule OPEN ACCESS molecules ISSN 1420-3049 /journal/molecules Article A Screening of a Library of T7 Phage-Displayed Peptide Identifies E2F-4 as an Etoposide-Binding Protein Mihoko Takami, Yoichi Takakusagi, Kouji Kuramochi, Senko Tsukuda, Satoko Aoki, Kengo Morohashi, Keisuke Ohta, Susumu Kobayashi, Kengo Sakaguchi and Fumio Sugawara * Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641, Yamazaki, Noda, Chiba 278-8510, Japan * Author to whom correspondence should be addressed; E-Mail: sugawara@rs.noda.tus.ac.jp; Tel.: +81-4-7124-1501 ext 3400; Fax: +81-4-7123-9767. Received: 1 March 2011; in revised form: 22 April 2011 / Accepted: 16 May 2011 / Published: 24 May 2011 Abstract: Etoposide (VP-16) is an anti-tumor compound that targets topoisomerase II (top II). In this study, we have identified an alternative binding protein of etoposide by screening a library of T7 phage-displayed peptides. After four rounds of selection using a biotinylated etoposide derivative immobilized on a streptavidin-coated plate, T7 phage particles that display a 16-mer peptide NSSASSRGNSSSNSVY (ETBP16) or a 10-mer NSLRKYSKLK (ETBP10) were enriched with the ratio of 40 or 11 out of the 69 clones, respectively. Binding of etoposide to these peptides was confirmed by surface plasmon resonance (SPR) analysis, which showed ETBP16 and ETBP10