World Journal of
Pharmaceutical and Life Sciences

CLONING PROTEIN EXPRESSION AND CHARACTERIZATION OF HUMAN RAB39B[1] YAAMINI SUBRAMANIAN

Yaamini Subramanian*

ABSTRACT

The Rab39b gene encodes a member of the Rab family of proteins. They are small GTPases that are involved in vesicular trafficking. In Rab proteins, the hydrolysis of GTP to GDP is coupled with association with and dissociation from membranes and it is present in all compartments of the endomembrane system (endoplasmic reticulum, Golgi, endosomes, lysosomes), the nucleus, the plasma membrane (including cell junctions and focal adhesions), mitochondria and centrioles. Our focus is to understand the structure and interactions of Rab39b protein using X-ray crystallography. Specifically we can study how proteins interact with other molecules, how they undergo conformational changes, and perform catalysis in the case of enzymes. Currently very little is known about Rab39B in the body and how loss of Rab39B contributes development of neurodegenerative disorder such as Parkinson’s disease, autism and Waisman syndrome. A codon optimized and the disordered regions at the C-terminal, removed Rab39b gene was cloned, fused with a glutathione S-transferase (GST) tag and sequenced. The cloned product was expressed in BL21 (DE3) cells resulting in a protein with molecular weight 19.8+25 kDa and verified by SDS-PAGE gel. Even though the function of Rab39b is less known, the protein has been implied to play important roles in synapse formation.

[Full Text Article]   [Download Certificate]