Drug receptor identification from multiple tissues using cellular-derived mRNA display libraries.
The use of display technologies to identify small molecule receptors from proteome libraries would provide a significant advantage in drug discovery. We have used mRNA display to select, based on affinity, proteins that bind to a drug of interest. A library of mRNA-protein fusion molecules was constructed from human liver, kidney, and bone marrow transcripts and selected using an immobilized FK506-biotin conjugate. Three rounds of selection produced full-length FKBP12 (FK506 binding protein 12 kDa) as the dominant clone. An analogous method was also used to map the minimal drug binding domain within FKBP12. Using this approach, it is anticipated that mRNA display could eventually play a key role in the discovery and characterization of new drug receptor interactions.
McPherson
Michael
M
Phylos, Inc., 128 Spring Street, Lexington, MA 02421, USA. mmcpherson@phylos.com
Yang
Yingfei
Y
Hammond
Philip W
PW
Kreider
Brent L
BL
D000595:Amino Acid Sequence; D001710:Biotin / Q000737:chemistry; D019921:Biotinylation; D001853:Bone Marrow / Q000378:metabolism; D003001:Cloning, Molecular; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D016559:Tacrolimus / Q000737:chemistry / Q000378:metabolism*; D022061:Tacrolimus Binding Protein 1A / Q000302:isolation & purification* / Q000378:metabolism
D019151:Peptide Library; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D001710:Biotin; D022061:Tacrolimus Binding Protein 1A; D016559:Tacrolimus