title: Human HOXA5 homeodomain enhances protein transduction and its application to vascular inflammation.
abstract: Cellular protein delivery is an emerging technique by which exogenous recombinant proteins are delivered into mammalian cells across the membrane. We have developed an Escherichia coli expression vector including human specific gene sequences for protein cellular delivery. The plasmid was generated by ligation the nucleotides 770-817 of the homeobox A5 mRNA sequence which was matched with protein transduction domain (PTD) of homeodomain protein A5 (HOXA5) into pET expression vector. The cellular uptake of HOXA5-PTD-EGFP was detected in 1min and its transduction reached a maximum at 1h within cell lysates. The cellular uptake of HOXA5-EGFP at 37°C was greater than in 4°C. For study for the functional role of human HOXA5-PTD, we purified HOXA5-APE1/Ref-1 and applied it on monocyte adhesion. Pretreatment with HOXA5-APE1/Ref-1 (100nM) inhibited TNF-α-induced monocyte adhesion to endothelial cells, compared with HOXA5-EGFP. Taken together, our data suggested that human HOXA5-PTD vector provides a powerful research tools for uncovering cellular functions of proteins or for the generation of human PTD-containing proteins.
authors.affiliation: Infectious Signaling Network Research Center and Research Institute for Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon 301-747, Republic of Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Lee
authors.forename: Kyoung Sook
authors.initials: KS
authors.lastname: Park
authors.forename: Eun Jung
authors.initials: EJ
authors.lastname: Cho
authors.forename: Hee Kyoung
authors.initials: HK
authors.lastname: Joo
authors.forename: Sang Ki
authors.initials: SK
authors.lastname: Lee
authors.forename: Sang Do
authors.initials: SD
authors.lastname: Lee
authors.forename: Jin Bong
authors.initials: JB
authors.lastname: Park
authors.forename: Seok Jong
authors.initials: SJ
authors.lastname: Chang
authors.forename: Byeong Hwa
authors.initials: BH
authors.lastname: Jeon
mesh_terms: D000595:Amino Acid Sequence; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000494:pharmacology*; D014657:Vasculitis / Q000378:metabolism*
chemical_list: C489213:HOXA5 protein, human; C560909:HOXA5-PTD-APE1-Ref-1 fusion protein, human; D018398:Homeodomain Proteins; D011993:Recombinant Fusion Proteins