![handle is hein.journals/injlegame106 and id is 1 raw text is: Int J Leg Med (1993) 106: 1-4
International Journal of
Legal Medicine
© Springer-Verlag 1993
Original articles
Human erythrocyte band 3 (EPB3) polymorphism:
analysis of blood and bloodstains by an immunodetection method
and frequency of the EPB3* Memphis variant
Akihiko Kimura, Tomoji Uda, Shoichi Nakashima, Haruhiko Ikeda, Seiji Yasuda, Motoki Osawa, and Tsutomu Tsuji
Department of Legal Medicine, Wakayama Medical College, 27 Kyuban-cho, 640 Wakayama, Japan
Received November 18, 1992 / Received in revised form March 24, 1993
Summary. The erythrocyte band 3 (EPB3) variant, band
3 Memphis (EPB3*Memphis), was detected by immuno-
blotting with a monoclonal antibody to the 41 kDa cyto-
plasmic N-terminal domain of band 3 without protease
treatment of erythrocytes. EPB3*Memphis was also de-
tected by immunoblotting from 3-month-old bloodstains
subjected to a-chymotrypsin treatment. A population
genetic study using this method indicated that the EPB3
variant would be useful for forensic work in Japan, since
the frequency of this variant in Japanese (Wakayama
prefecture) is relatively high (0.159).
Key words: Band 3 Memphis - Erythrocyte band 3 - Ge-
netic polymorphism - Monoclonal antibody
Zusammenfassung. Eine Variante der Erythrozyten-
Bande 3 (EPB 3), namlich die Memphis-Variante (EPB
3*Memphis), wurde mit Hilfe eines monoklonalen Anti-
korpers, welcher gegen die 41 kDa zytoplasmatische N-
terminale Domane der Bande 3 gerichtet ist, und des
Immunoblottings ohne Proteasebehandlung der Erythro-
zyten nachgewiesen. EPB 3*Memphis wurde auch mit
Hilfe des Immunoblottings aus 3 Monate alten Blutspu-
ren nachgewiesen, welche einer a-Chymotrypsinbehand-
lung unterzogen wurden. Eine populationsgenetische
Studie mit Hilfe dieser Methode zeigte, daB die EPB*3-
Variante nutzlich fur forensische Arbeiten in Japan sein
wurde, da die Haufigkeit dieser Variante bei Japanern
(Prafektur Wakayama) relativ hoch ist (0,159).
Schlisselworter: Bande 3-Memphis - Erythrozyten-
Bande 3 - Genetischer Polymorphismus - Monoklonaler
Antikorper
Introduction
Erythrocyte band 3, a transmembrane glycoprotein (Mr
= 90-100 kDa) which acts as an anion exchanger and
Correspondence to: A. Kimura
plays a key role in membrane stability, is one of the major
components of the human erythrocyte membrane [1, 2].
The C-terminal domain of band 3 (Mr = 55-60 kDa)
spans the membrane several times and is a functional
domain for anion exchange. The cytoplasmic N-terminal
domain of band 3 (Mr = 41 kDa), however, binds to
cytoskeletal proteins and glycolytic enzymes, and is not
essential for anion exchange [3, 4]. Band 3 is detected as
a diffuse band in sodium dodecyl sulfate-polyacrylamide
gel electrophoresis (SDS-PAGE), since a highly hetero-
genous carbohydrate chain is linked to the asparagine
residue at its C-terminal domain. Band 3 is polymorphic
and several variants have been reported [5-9]. The locus
for band 3 (EPB 3) has been assigned to chromosome
17q12-q21 [10] and EPB 3*Memphis [11] is a genetically
determined variant at the EPB 3 locus. The alternative
allele is inherited codominantly and has been found to
occur at appreciable frequencies in all populations
tested, the frequency being higher in Asians and Afri-
cans than in Caucasians [11]. No abnormalities in the
function or shape of erythrocytes have been observed
even in homozygotes [5]. The amino acid substitution
(Lys 56-+ Glu) responsible for this variant gives N-termi-
nal proteolytic fragments with Mr = 63 kDa and 43 kDa
instead of those with Mr = 60 kDa and 41 kDa found in
normal band 3 by SDS-PAGE [12]. This variant can be
detected only in the proteolytic fragments of band 3 and
not in intact band 3 by SDS-PAGE, since it is masked by
the highly heterogeneous carbohydrate chain [5].
In this paper, we describe the immunological detec-
tion of EPB 3*Memphis from erythrocytes and blood-
stains, and we report the gene frequency for this variant
in a Japanese population.
Materials and methods
Specimens. Blood samples were collected from healthy donors and
prepared immediately. Outdated blood samples, used for the
population genetic study of EPB 3 polymorphism, were obtained
from the Wakayama Red Cross Blood Center. Bloodstains were](ViewImageLocal?handle=hein.journals/injlegame106&div=1&collection=&method=preview&ext=.png&size=3)
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