TY  - JOUR
TI  - KDEL tagging: a method for generating dominant-negative inhibitors of the secretion of TGF-beta superfamily proteins
AU  - Matsukawa, Shinya
AU  - Moriyama, Yuki
AU  - Hayata, Tadayoshi
AU  - Sasaki, Haruka
AU  - Ito, Yuzuru
AU  - Asashima, Makoto
AU  - Kuroda, Hiroki
T2  - The International Journal of Developmental Biology
AB  - Most endoplasmic reticulum (ER)-retained proteins contain a carboxy-terminal signal sequence called the ER retention signal motif such as the Lys-Asp-Glu-Leu (KDEL) motif. Using this molecular mechanism, we developed a new dominant-negative assay, designated the KDEL-tag trap assay, to negatively regulate secretion of disulfide bond-dependent protein dimers, as typified by TGF-beta superfamily proteins. First, we tested this method on the Nodal protein Xnr5, which is a well-studied mesoderm inducer in vertebrates. Tagging of Xnr5 protein with KDEL at the carboxy-terminus effectively blocked the secretion of Xnr5, resulting in complete inhibition of mesoderm induction in Xenopus embryogenesis. Second, we examined the usefulness of the KDEL-tag trap assay on BMPs, which are well-known negative regulators of neural induction and ventralizing factors during early development, and demonstrated that the functions of the BMP family proteins BMP4 and ADMP were blocked by the KDEL-tag trap assay. Moreover, the technical feasibility of the KDEL-tag trap assay was confirmed in a cell culture system using mouse osteoblasts. Taken together, these results suggest that the KDEL-tag trap assay can be adapted to inhibit a variety of plasma membrane or secreted proteins of a multimeric nature.
PY  - 2012
DO  - 10.1387/ijdb.123514sm
VL  - 56
IS  - 5
SP  - 351
EP  - 356
J2  - Int. J. Dev. Biol.
LA  - en
SN  - 0214-6282
SN  - 1696-3547
UR  - https://ijdb.ehu.eus/article/123514sm
Y2  - 2024/05/02/17:51:12
ER  -