What might cause a recombinant protein not to be secreted to the extracellular matrix otherwise getting stucked in the cell. This is a domain of a recombinatly expressed extracellular protein and everything seems to be fine with the signal peptide sequence and cells looks healthy
Need ECM remodelers to activate secretion from ECM (not secretion into ECM). Usually MMPs.
Please can you explain more. my protein is an extracellular protein and it is suppose to be secreted outside the cell into the supernatant under normal circumstances but instead it is retain in the cell lysate
Most protein have latency associated peptide which holds the proteins at ECM through protein protein/carbohydrate interactions. When the activation signal comes, either it lose interaction or cleaved to release active secretory protein. This cleavage is usually made by matrixmetalloproteinases (MMPs). So it is likely that it is not cleaved, so still stuck to ECM and hence you see it in cell lysate.
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