Study on the function of Rh glycoprotein in rainbow trout oocytes using non-invasive micro-test

Amine transmembrane transport is a key factor in maintaining nitrogen metabolism and nitrogen balance in organisms. Rainbow trout gill amine transport may be regulated by Rh glycoprotein. In 2010, Canadian scientists used non-invasive micro-testing techniques to study the amine transport of different rainbow trout glycoproteins (R Rhag, Rhbg1, Rhbg2, Rhcg1, Rhcg2, Rh30-like) expressed in oocytes. Except for Rh30-like, other Rh proteins can promote the absorption of methylamine, the absorption can reach saturation, and the Km value is 4.6-8.9 mM. The extracellular pH increased from 7.5 to 8.5, resulting in a 3-4 fold increase in the maximum flow rate (Jmax) of methylamine. The Km value did not change when all glycoproteins were expressed or protonated methylamine. The ratio of the pH gradient stimulating the outflow and inflow of methylamine is positively correlated with the increase in the H + gradient. This study confirmed that Rhag and Rhcg2 promote the absorption of amines, but Rh30-like protein cannot promote the absorption using non-invasive micro-testing technology. At low substrate concentrations, the absorption rate of amine is higher than that of methylamine. The Km value of the amine is within the physiological range of rainbow trout. This study found that the Rh glycoprotein of rainbow trout gills has a low affinity and a high amine transport ability to form a pH gradient. When the concentration of cytosolic amines increases, the rapid outflow of amines is promoted. This study provides direct evidence for our understanding of the transport of amines. The non-invasive micro-measurement technique can directly measure the flow rate of NH4 + and NO3- in vivo, which is of great significance for studying the transport of nitrogen and the function of proteins. Keywords: amine transport, Rh glycoprotein, oocyte, rainbow trout, non-invasive micro-testing technique References: Nawata CM, et al. Journal of Experimental Biology, 2010: 213, 1049-1059.

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