Process for heterologously expressing and purifying human glucose transporters GLUT1, GLUT2 and GLUT3

Abstract

The invention discloses a method for heterologous expression and purification of members of glucose transporter family including CLUT1, CLUT2, and CLUT3 on human cell membrane, which comprises the following steps of: constructing an inducible fusion protein expression vectors of GST-CLUT1, GST-GLUT2, and GST-GLUT3 and transforming fission yeast; identifying transformants of the fission yeast; induced expression of the three fusion proteins of GST-CLUT1, GST-GLUT2, and GST-GLUT3 in the fission yeast; identifying the expression of the genes of GLUT1, GLUT2 and GLUT3 in the fission yeast; separating and purifying the three fusion protein; detecting and identifying the three separated and purified fusion proteins; separating and purifying human proteins of GLUT1, GLUT2 and GLUT3 from the three fusion proteins respectively.

Description

Method for heterologous expression and purification of human glucose transporter family members GLUT1, GLUT2, GLUT3 technical field The invention relates to a method for expressing and purifying proteins, in particular to a method for heterologously expressing and purifying glucose transporter family members GLUT1, GLUT2 and GLUT3 (facilitated glucose transporter, GLUTs) on human cell membranes. Background technique Glucose transporters (facilitatedglucosetransporter, GLUTs) located on the human cell membrane are an important class of membrane proteins in the body. They promote the transmembrane transport of glucose in the body and play a vital role in stabilizing cell energy and maintaining life activities. So far, a variety of GLUTs have been discovered and identified. They are encoded by different genes, have similar protein structures and biological functions, and their gene expression and intracellular distribution are regulated by many factors. Human GLUTs are compos...

AI Technical Summary

Problems solved by technology

However, like most membrane proteins, the structural characteristics of GLUTs determine that there are many difficulties in expression, isolation, and purification.

The main reasons are: (1) the content of natural membrane proteins is low, most of which are lower than the order of micrograms; (2) membrane proteins are difficult to express in vitro, and there has been no report on a high-efficiency expression system for membrane proteins so far; (3) membrane proteins are unstable and difficult to separate , Purification is more difficult

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