์ฝ˜ํ…์ธ ๋กœ ๊ฑด๋„ˆ๋›ฐ๊ธฐ
Merck

Microvesicle-mediated release of soluble LH/hCG receptor (LHCGR) from transfected cells and placenta explants.

Reproductive biology and endocrinology : RB&E (2011-05-18)
Anne E Chambers, Paul F Stanley, Harpal Randeva, Subhasis Banerjee
์ดˆ๋ก

Placental hCG and pitutary LH transduce signals in target tissues through a common receptor (LHCGR). We demonstrate that recombinant LHCGR proteins which include the hormone-binding domain are secreted from transfected cells and that natural LHCGR is also secreted from human placental explants. LHCGR recombinant proteins representing varying lengths of the N-terminal extracellular domain were expressed in Chinese Hamster Ovary cells in suspension culture. Secretion was minimal up to 72h but by 96h 24-37% of the LHCGR had been released into the culture medium. The secreted proteins were folded and sensitive to glycosidases suggesting N-linked glycosylation. Secretion was independent of recombinant size and was mediated via structurally defined membrane vesicles (50-150nm). Similarly cultured human early pregnancy placental explants also released LHCGR via microvesicles. These studies provide the first experimental evidence of the possible mechanistic basis of the secretion of LHCGR.

MATERIALS
์ œํ’ˆ ๋ฒˆํ˜ธ
๋ธŒ๋žœ๋“œ
์ œํ’ˆ ์„ค๋ช…

Sigma-Aldrich
Monoclonal ANTI-FLAGยฎ antibody produced in rabbit, clone SIG1-25, ascites fluid
Sigma-Aldrich
ANTI-FLAGยฎ antibody, Rat monoclonal, clone 6F7, purified from hybridoma cell culture
Sigma-Aldrich
IGEPALยฎ CA-630, for molecular biology
Sigma-Aldrich
IGEPALยฎ CA-630, viscous liquid
Supelco
IGEPALยฎ CA-630
Cap for Flask