Stroke, or cerebral apoplexy, is a paroxysmal disease that seriously threatens the neurological functions of middle-aged and older men, resulting in high rates of disability and mortality. The blood level of Corin can accurately predict the risk and subtype of stroke. Herein, an immunosensor was developed utilizing a nanocomposite of Au nanoparticles and Co 3 O 4 nanocubic cages (AuNPs/Co 3 O 4 NcC) as the sensing substrate, with luminol serving as the sensing probe for Corin detection. Through the application of 3-aminopropyltrimethoxysilane, AuNPs/Co 3 O 4 NcC was affixed to the surface of indium tin oxide (ITO)-coated glass, considerably enhancing the electrochemiluminescence (ECL) performance of luminol due to its peroxidase-like activity. To confer biological specificity by immobilizing the Corin antibody on this substrate and to block residual active sites with bovine serum albumin, an immunosensor was developed. The capture of Corin protein by the antibody through an immune reaction, leading to its binding on the sensor, resulted in a decreased ECL signal. Under optimized conditions, the proposed ECL immunosensor demonstrated a broad linear response range from 1 pg/mL to 4800 pg/mL and a low detection limit of 0.26 pg/mL. This ECL immunosensor is characterized by its specificity, stability, and reproducibility and can detect Corin in real blood samples.
Selective protein distribution on distinct plasma membranes is important for epithelial cell function. To date, how proteins are directed to specific epithelial cell surface is not fully understood. Here we report a conserved DSSDE motif in LDL-receptor (LDLR) modules of corin (a transmembrane serine protease) and CD320 (a receptor for vitamin B12 uptake), which regulates apical membrane targeting in renal epithelial cells. Altering this motif prevents specific apical corin and CD320 expression in polarized Madin–Darby canine kidney (MDCK) cells. Mechanistic studies indicate that this DSSDE motif participates in a Rab11a-dependent mechanism that specifies apical sorting. In MDCK cells, inhibition of Rab11a, but not Rab11b, expression leads to corin and CD320 expression on both apical and basolateral membranes. Together, our results reveal a novel molecular recognition mechanism that regulates LDLR module-containing proteins in their specific apical expression in polarized renal epithelial cells.