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Use of domain-deletion mutants to locate Lutheran blood group antigens to
each of the five immunoglobulin superfamily domains of the Lutheran
glycoprotein: elucidation of the molecular basis of the Lu(a)/Lu(b) and the
Au(a)/Au(b) polymorphisms
SF Parsons, G Mallinson, GL Daniels, CA Green, JS Smythe and DJ Anstee
International Blood Group Reference Laboratory, Bristol Institute for
Transfusion Sciences, UK.
Lutheran glycoprotein (Lu gp) has five predicted immunoglobulin superfamily
(IgSF) domains. K562 cells were transfected with Lu cDNA and tested by flow
cytometry with monoclonal antibodies and Lu blood group antisera. The
results confirmed the identity of Lu cDNA. Deletion mutants lacking the
regions encoding one or more IgSF domains were made by inverse polymerase
chain reaction (PCR), expressed in K562 cells, and tested with the same
antibodies. The Lu(b) and Lu5 antigens and the epitope recognized by
monoclonal antibody BRIC 224 were mapped to the first, N-terminal, IgSF
domain. Lu4 and Lu8 were mapped to domain 2; Lu20 to domain 3; Lu7 and BRIC
221 epitope to domain 4, and Lu13 and Au(b) to domain 5. The organization
of the LU gene was determined. The region encoding the open reading frame
is arranged in 15 exons extending over approximately 11 kb on chromosome
19q13.2. The Lu(a)/Lu(b) and Au(a)/Au(b) blood group polymorphisms were
studied using genomic DNA from typed blood donors. The Lu(a) mutation is a
base change in exon 3 (G252 to A) encoding an Arg77 (Lu(b)) to His (Lu(a))
change on the CFG face of domain 1. The Au(a)/Au(b) polymorphism is an
A1637 to G substitution in exon 12 encoding a Thr539 (Au(a)) to Ala (Au(b))
change on the G strand of domain 5.
Volume 89,
Issue 11,
pp. 4219-4225,
06/01/1997
Copyright © 1997 by The American Society of Hematology
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