HSPA8, not C3AR1 trapped from SH-SY5Y cells: demonstrated by TLQP-21 avidin agarose affinity chromatography HSPA8, not C3AR1 trapped by TLQP-21 in SH-SY5Y cells

Abstract

Md. Shamim Akhter , Jesus Rodriguez Requena

To ‘fish out’ HSPA8 as a receptor of human TLQP-21 in human neuroblastoma SH-SY5Y cell; cross-linking, affinity chromatography and mass spectrometry-based protein identification techniques were used (by our group). Before that, in rodent cells, C3AR1 was found as a receptor of rodent TLQP-21 in CHO-K1 cells (by another group). So, it was of very much interest to find out whether C3AR1 can be ‘fished out’ as a receptor of human TLQP-21 in a human SH-SY5Y cell line or not. Here PVDF membrane (which was used for immunohistochemical validation of HSPA8 in SH-SY5Y cell line) was stripped, followed by immunohistochemical validation as before under the same condition, but C3AR1 was not found to bind with human TLQP-21. The findings concluded here that in SH-SY5Y cells, HSPA8 but not C3AR1 was documented as a receptor of human TLQP-21 using avidin agarose affinity chromatography. The significance of the study is that it provides a starting point to signpost: human TLQP-21 exerts its biological activity via HSPA8 (not C3AR1) in human SH-SY5Y cells. The further readout of TLQP-21-HSPA8 signaling can be exploited to explore new horizon in diagnosis and therapies for VGF related human diseases, especially in which TLQP-21 has been shown to affect and related with thereof.

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