Characterization of rhodamine conjugated Agiotensin II peptide: synthesis, analysis and receptor binding and internalization.
The results in this study show that the rhodamine fluorophore can be specifically conjugated to Angiotensin II at Lys3 residue (substituted for a Val) without altering the biological activity of the parent compound. The conjugated peptide was characterized using HPLC, mass spectrometry, and N-terminal sequencing. The rhodamine-Angiotensin II binds effectively to AT1 receptor and gets internalized in clathrin coated vesicles by endocytosis. These results clearly suggest the usefulness of fluorophore-conjugated peptides in studies such as, ligand-receptor binding, and ligand-receptor complex internalization, for drug delivery using cell receptors and as an alternative to peptide hormone radioimmunoassays.
Yadav
Satya P
SP
Biotechnology Biotechnology Core, The Lerner Research Institute, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, Ohio 44195, USA. yadavs@ccf.org
Karnick
Sadashiv
S
Shen
Wei-Zhen
WZ
Zhang
Jingli
J
D000804:Angiotensin II / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D007295:Inositol Phosphates / Q000096:biosynthesis; D013058:Mass Spectrometry; D011945:Receptors, Angiotensin / Q000378:metabolism; D012235:Rhodamines / Q000737:chemistry / Q000378:metabolism*; D013194:Staining and Labeling
D007295:Inositol Phosphates; D011945:Receptors, Angiotensin; D012235:Rhodamines; D000804:Angiotensin II