Defining the Rh blood group antigens. Biochemistry and molecular genetics

The Rh blood group antigens (D, Cc and Ee series) are carried by a family of non glycosylated hydrophobic transmembrane proteins of 30-32 kDa which are missing from the red cells of rare Rhnull individuals that express several membrane defects. The structure of these proteins has been deduced from cDNA cloning and studies have shown that the Rh proteins are erythroid specific and share no sequence homology with any known protein. The RhD and non-D proteins exhibit 92% sequence identity and their predicted membrane topology is similar as most of the molecules appear to reside between the leaflets of the phospholipid bilayer with only short hydrophilic loops connecting the twelve putative transmembrane helices. The RHD and RHCE genes encoding the Rh proteins (D and Cc/Ee, respectively) are organized in tandem on chromosome 1p34-p36 and most likely derived by duplication of a common ancestral gene. This concept is supported by the identification of RH-like genes in non human primates. The human RH locus is best described as a two-gene model in which all RhD-positive and most RhD-negative haplotypes are composed of two (RHD and RHCE) or only one (RHCE) structural genes, respectively. The RHD gene encodes the D protein and the RHCE gene encodes the C/c and E/e proteins presumably by alternative splicing of a pre messenger RNA. The correlation between the blood group D epitopes and the amino acid polymorphism of the Rh proteins is not yet established, but amino acid polymorphisms at positions 103 and 226 determine the molecular basis for the C/c (Ser-->Pro) and E/e (Pro-->Ala) specificities, respectively. Most variants analyzed so far are caused by gene conversion which appears as the principal mechanism responsible for polymorphism and gene diversity in the RH system. However, gene deletions have also been found in some occasions. To date, all Rhnull phenotypes investigated most likely result from transcriptional regulatory mechanisms that are not yet understood. Rhnull individuals suffer a clinical syndrome of varying severity and their red cells are characterized by morphological and functional abnormalities of cation transport and phospholipid asymmetry. In addition, several membrane components including the Rh proteins and other glycoproteins recently characterized (Rh50 glycoprotein, CD47, glycophorin B, Duffy, LW) are absent or severely decreased on these cells. These findings suggest that the Rh proteins are assembled into a multimeric complex with these glycoproteins and further studies should clarify the role in biosynthesis and the potential function of each component in this complex.