Producing recombinant human mucosal addressin cell adhesion molecule 1 (MADCAM1) in yeast involves co-cloning the gene of interest (19-317aa of human MADCAM1) into an expression vector with a C-terminal 6xHis-tag gene, which is transformed into yeast cells. The cells are cultured to induce protein expression. After reaching adequate growth, the cells are lysed to extract the recombinant MADCAM1 protein. The collected proteins are purified through affinity chromatography. The purity of the recombinant MADCAM1 protein is assessed using SDS-PAGE, exceeding 85%.
Human MADCAM1 is expressed by fibroblastic reticular cells (FRCs) in the lymph node microenvironment and plays a crucial role in leukocyte trafficking to the gut-associated lymphoid tissues (GALTs) [1]. It is a ligand for α4β7 integrin, facilitating the recruitment of lymphocytes to the gut mucosa [2]. The expression of MADCAM1 is equally distributed throughout the proximal and distal colon, contributing to the homing of dendritic cells to the human colon [2].
Furthermore, MADCAM1 is involved in the selective recruitment of peripheral blood CD16(–) natural killer cells into the human endometrium [3]. It interacts with L-selectin and chondroitin sulfate proteoglycans, mediating the migration of specific immune cells to the endometrial tissue [3].
References:
[1] A. Habtezion, L. Nguyen, H. Hadeiba, & E. Butcher, Leukocyte trafficking to the small intestine and colon, Gastroenterology, vol. 150, no. 2, p. 340-354, 2016. https://doi.org/10.1053/j.gastro.2015.10.046
[2] D. Bernardo, L. Durant, E. Mann, E. Bassity, E. Montalvillo, R. Manet al., Chemokine (c-c motif) receptor 2 mediates dendritic cell recruitment to the human colon but is not responsible for differences observed in dendritic cell subsets, phenotype, and function between the proximal and distal colon, Cellular and Molecular Gastroenterology and Hepatology, vol. 2, no. 1, p. 22-39.e5, 2016. https://doi.org/10.1016/j.jcmgh.2015.08.006
[3] T. Yamaguchi, K. Kitaya, N. Daikoku, T. Yasuo, S. Fushiki, & H. Honjo, Potential selectin l ligands involved in selective recruitment of peripheral blood cd16(–) natural killer cells into human endometrium1, Biology of Reproduction, vol. 74, no. 1, p. 35-40, 2006. https://doi.org/10.1095/biolreprod.105.045971
