Journal of Glycomics & Lipidomics
Open Access

Abstract

Label free quantitative proteomics approach unravels the pleiotropy of buffalo leukemia inhibitory factor (BuLIF) in COS-1 cells

Sudarshan Kumar

Leukemia inhibitory factor (LIF) is a pleotropic molecule which performs diverse functions in a context dependent manner. Bovine LIF (BuLIF) is an essential media component in in-vitro stem cell culture and also considered essential in the early stages of pregnancy. However, the exact molecular mechanism behind the diverse actions of this molecule is unknown except the stat3 mediated canonical pathways in stem cell pluripotency. We produced a stably transfected COS-1_BuLIF cell line which expressed high amount of LIF in media. The integration of BuLIF into genome was confirmed by PCR followed by sequencing. We found that LIF induces dome like structure formation which is indicative of BuLIF action via stat3 pathway. Further, pure rBuLIF was purified from this cell line which was found to be 58.99 kDa and 48.9 kDa protein with and without glycosylation respectively which was confirmed by western blot and nLC-MS/MS. The time lapse and concentration-dependent assay of purified LIF showed maximum inhibition at 72 hours and half-maximal effective concentration (EC50) to be 0.0555 ng/mL, corresponding to a specific activity of >1.6

Published Date: 2021-08-15;