Xenbase Image ID: 144724

Figure 4. Western blot analysis of bovine wild-type rhodopsin and bovine P23H rhodopsin from transgenic X. laevis eyes indicate that bovine P23H rhodopsin exists primarily in an N-terminally truncated form and is more abundant in dark-reared retinas. A, Western blot analysis was performed using samples of bovine rhodopsin deglycosylated with PNGaseF (a, e) and extracts from transgenic X. laevis eyes expressing bovine wild-type rhodopsin in cyclic light (b, f), bovine P23H rhodopsin in cyclic light (c, g), and bovine P23H rhodopsin in constant dark (d, h). Western blots were probed with mAb 1D4 (recognizing the C terminus; a–d) or mAb A5-3 (recognizing the N terminus; e–h). Retinas expressing bovine P23H rhodopsin contained large quantities of a truncated rhodopsin that was nonreactive with mAb A5-3. B, Western blot of extracts of retinas expressing bovine P23H rhodopsin (a) and N-terminal truncated rhodopsin (ρΔ2–23) (b) demonstrating that the truncated form of bovine P23H rhodopsin is missing at least 23 amino acids from the N terminus. C, Western blot of extracts of retinas expressing bovine wild-type reared in cyclic light (a) and P23H/K296R rhodopsin reared in darkness (b) or cyclic light (c). P23H/K296R rhodopsin is truncated in both dark and cyclic light conditions. D, Sequence alignment of the N-terminal 35 amino acid residues of X. laevis, bovine, human, and mouse rhodopsin transgenes. Amino acid positions that vary among species are indicated by asterisks, and the region encompassing the putative cleavage site is underlined. The P23 residue is indicated by an arrow. Image published in: Tam BM and Moritz OL (2007) Copyright © 2007. This image is reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

Return to previous page