Immunochemical analysis of the Kx protein from human red cells of different Kell phenotypes using antibodies raised against synthetic peptides

Carbonnet, Florence; Hattab, Claude; Collec, Emmanuel; Kim, Caroline Le Van; Cartron, Jean‐Pierre; Bertrand, Olivier

doi:10.1046/j.1365-2141.1997.d01-2110.x

Cited by 41 publications

(24 citation statements)

References 19 publications

(44 reference statements)

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“…Nevertheless, the cellular expression of Kell and XK seems to be interconnected as absence of XK in McLeod muscle is accompanied by a marked decrease in Kell, to an extent that Kell was not detectable by immunostaining. This relation between Kell and XK expression in muscle is comparable to the situation in Kell‐null red blood cells, which lack Kell protein and have a diminished amount of XK protein 6. Our results showing different cellular localizations for Kell and XK suggest independent functions for each of the proteins in the muscle.…”

Section: Discussionsupporting

confidence: 79%

Kell and XK immunohistochemistry in McLeod myopathy

Jung¹,

Russo²,

Redman³

et al. 2001

Muscle and Nerve

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The McLeod syndrome is an X-linked neuroacanthocytosis manifesting with myopathy and progressive chorea. It is caused by mutations of the XK gene encoding the XK protein, a putative membrane transport protein of yet unknown function. In erythroid tissues, XK forms a functional complex with the Kell glycoprotein. Here, we present an immunohistochemical study in skeletal muscle of normal controls and a McLeod patient with a XK gene point mutation (C977T) using affinity-purified antibodies against XK and Kell proteins. Histological examination of the affected muscle revealed the typical pattern of McLeod myopathy including type 2 fiber atrophy. In control muscles, Kell immunohistochemistry stained sarcoplasmic membranes. XK immunohistochemistry resulted in a type 2 fiber-specific intracellular staining that was most probably confined to the sarcoplasmic reticulum. In contrast, there was only a weak background signal without a specific staining pattern for XK and Kell in the McLeod muscle. Our results demonstrate that the lack of physiological XK expression correlates to the type 2 fiber atrophy in McLeod myopathy, and suggest that the XK protein represents a crucial factor for the maintenance of normal muscle structure and function.

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Section: Discussionsupporting

confidence: 79%

Kell and XK immunohistochemistry in McLeod myopathy

Jung¹,

Russo²,

Redman³

et al. 2001

Muscle and Nerve

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“…Polyclonal antibodies (PAbs): The rabbit antibodies raised against the 43 kDa N-terminal cytoplasmic domain of human Band 3 and against protein 4.1R (el Ouggouti , et al 1992) were generous gifts of Dr D. Dhermy (INSERM, U665, Paris, France); anti-4.2 was described previously (Hayette , et al 1995, Jons and Drenckhahn 1992); MPC8 raised against the C-terminal region of the Rh polypeptides was described previously (Hermand , et al 1993); rabbit anti-Kx2 obtained by immunization against a synthetic peptide containing XK residues 111 to 127 located in the large extracellular loop of the protein XK (Carbonnet , et al 1997); anti- aquaporin (AQP1) antibody (AB3065, Chemicon, Billerica, MA, USA); anti-Co3 (Sar serum) was from the CNRGS (Institut National de la Transfusion Sanguine, Paris, France). For UT-B, the antibody was raised against a 20-amino acid peptide corresponding to the C-terminal sequence of rat UT-B (Trinh-Trang-Tan , et al 2002).…”

Section: Methodsmentioning

confidence: 99%

The human Kell blood group binds the erythroid 4.1R protein: new insights into the 4.1R-dependent red cell membrane complex

et al. 2015

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Summary Protein 4.1R plays an important role in maintaining the mechanical properties of the erythrocyte membrane. We analysed the expression of Kell blood group protein in erythrocytes from a patient with hereditary elliptocytosis associated with complete 4.1R deficiency (4.1(−) HE). Flow cytometry and Western blot analyses revealed a severe reduction of Kell. In vitro pull down and co-immunoprecipitation experiments from erythrocyte membranes showed a direct interaction between Kell and 4.1R. Using different recombinant domains of 4.1R and the cytoplasmic domain of Kell, we demonstrated that the R46R motif in the juxta-membrane region of Kell binds to lobe B of the 4.1R FERM domain. We also observed that 4.1R deficiency is associated with a reduction of XK and DARC (also termed ACKR1) proteins, the absence of the glycosylated form of the urea transporter B and a slight decrease of band 3. The functional alteration of the 4.1(−) HE erythrocyte membranes was also determined by measuring various transport activities. We documented a slower rate of HCO3−/Cl− exchange, but normal water and ammonia transport across erythrocyte membrane in the absence of 4.1. These findings provide novel insights into the structural organization of blood group antigen proteins into the 4.1R complex of the human red cell membrane.

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“…The XK protein is predicted to have 10 transmembrane domains with structural characteristics of a membrane transporter protein, but its function is yet to be defined. In MLS RBCs, truncation of the XK protein is associated with reduced expression of the Kell blood group antigen and a compensated hemolytic anemia [70][71][72][73]. 2) [29,68,69].…”

Section: Mcleod Syndromementioning

confidence: 99%