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Mutagenesis Services> | The hsc-70 mutants (R533A, R535A, K573Q, K583Q, K589Q, K597Q, and K601Q) were generated using site-directed mutagenesis with the mouse wild-type hsc-70 cDNA as template (GenScript, Piscataway, NJ). | Get A Quote |
hsc-70 (HSPA8) is a cytosolic molecular chaperone, which plays a central role in cellular proteostasis, including quality control during protein refolding and regulation of protein degradation. hsc-70 is pivotal to the process of macroautophagy, chaperone-mediated autophagy, and endosomal microautophagy. The latter requires hsc-70 interaction with negatively charged phosphatidylserine (PS) at the endosomal limiting membrane. Herein, by combining plasmon resonance, NMR spectroscopy, and amino acid mutagenesis, we mapped the C terminus of the hsc-70 LID domain as the structural interface interacting with endosomal PS, and we estimated an hsc-70/PS equilibrium dissociation constant of 4.7 ± 0.1 ... More