The production of recombinant mouse fatty acid-binding protein, liver protein (Fabp1) begins with isolating the target gene that codes for the 1-127aa of mouse Fabp1 protein. This gene is cloned into an expression vector and introduced into E. coli cells through transfection. The E. coli cells express the protein, which is subsequently harvested from the cell lysate. The protein is purified using affinity chromatography. Finally, the protein's functionality is validated through various biochemical assays to ensure it performs its intended function effectively. Its purity is over 95% as determined by SDS-PAGE. It contains less than 1.0 EU/μg of endotoxin as measured by the LAL method. It is an active protein. The binding affinity of this recombinant FABP1 for the synthetic ligand cis-parinaric acid, measured by fluorescence titration, shows that half-maximal fluorescence of 2.5 μM rMuFABP1 is achieved with approximately 5 μM cis-parinaric acid.
Mouse Fabp1 is primarily involved in fatty acid uptake, oxidation, and very low-density lipoprotein secretion in the liver [1]. Research indicates that Fabp1 acts as an antioxidant protein, helping to mitigate hepatic oxidative stress during chronic ethanol ingestion [2]. Fabp1 is unique in that it is the only isoform capable of binding both fatty acids and fatty acyl-CoA, setting it apart from other Fabp isoforms [3]. Fabp1 interacts with plasma membrane proteins like FATP5, influencing fatty acid translocation and uptake [4]. Studies have shown that Fabp1 expression is associated with hepatic lipid metabolism, with implications in conditions such as non-alcoholic fatty liver disease [5]. Fabp1 has been linked to hepatic steatosis and inflammation, impacting the biotransformation of substances like δ9-THC [6]. Furthermore, Fabp1 also regulates lipid metabolism in various tissues, including facilitating triacylglycerol synthesis and secretion in the liver [7].
References:
[1] T. Mukai, M. Egawa, T. Tamaki, H. Yamashita, & T. Kusudo, Silencing of fabp1 ameliorates hepatic steatosis, inflammation, and oxidative stress in mice with nonalcoholic fatty liver disease, Febs Open Bio, vol. 7, no. 7, p. 1009-1016, 2017. https://doi.org/10.1002/2211-5463.12240
[2] G. Wang, H. Bonkovsky, A. Lemos, & F. Burczynski, Recent insights into the biological functions of liver fatty acid binding protein 1, The Journal of Lipid Research, vol. 56, no. 12, p. 2238-2247, 2015. https://doi.org/10.1194/jlr.r056705
[3] T. Grevengoed, E. Klett, & R. Coleman, Acyl-coa metabolism and partitioning, Annual Review of Nutrition, vol. 34, no. 1, p. 1-30, 2014. https://doi.org/10.1146/annurev-nutr-071813-105541
[4] F. Schroeder, A. McIntosh, G. Martin, H. Huang, D. Landrock, S. Chunget al., Fatty acid binding protein‐1 (fabp1) and the human fabp1 t94a variant: roles in the endocannabinoid system and dyslipidemias, Lipids, vol. 51, no. 6, p. 655-676, 2016. https://doi.org/10.1007/s11745-016-4155-8
[5] H. You, H. Ma, X. Wang, X. Wen, C. Zhu, W. Maoet al., Association between liver-type fatty acid-binding protein and hyperuricemia before and after laparoscopic sleeve gastrectomy, Frontiers in Endocrinology, vol. 13, 2022. https://doi.org/10.3389/fendo.2022.993137
[6] M. Elmes, L. Prentis, L. McGoldrick, C. Giuliano, J. Sweeney, O. Josephet al., Fabp1 controls hepatic transport and biotransformation of δ9-thc, Scientific Reports, vol. 9, no. 1, 2019. https://doi.org/10.1038/s41598-019-44108-3
[7] Y. Wang, K. Tang, W. Zhang, W. Guo, Y. Wang, & Z. Liu, Fatty acid-binding protein 1 increases steer fat deposition by facilitating the synthesis and secretion of triacylglycerol in liver, Plos One, vol. 14, no. 4, p. e0214144, 2019. https://doi.org/10.1371/journal.pone.0214144
