Biochimica et Biophysica Acta, 1096 (1991) 263-264 263

~ 1991 Elsevier Science Publishers B.V. 0925-4439/91/$03.50

ADONIS 0925443991000733

BBADIS 60003 Rapid Report

Malaria and ovalocytosis - molecular mimicry?

A. Husain-Chishti and Paul Ruff
Malaria Research Laboratory, Department of Biomedical Research, St. Elizabeth's Hospital, Tufts University School of Medicine, Boston,
MA (U.S.A.)

(Received 29 January 1991)

Key words: Ovalocytosis; Malaria; (P. falciparum)

Two recently published reports have described findings which will have a profound impact on the understanding of
molecular mechanisms of human resistance to malaria infection. In Melanesian ovalocytosis, a genetic polymorphism
found in Papua New Guinea and parts of South East Asia, the red cells are highly resistant to invasion by various
species of malaria parasite. The molecular nature of the defect in ovalocytic erythrocytes was not known. Recent reports
by Liu et al. (Liu, S.-C., Zhai, S., Palek, J., Golan, D., Amato, D., Hassan, K., Nurse, G., Babona, D., Coetzer, T.,
Jarolim, P. Zaik, M. and Borwein, S. (1990) N. Engl. J. Med. 323, 1530-1538.) and Jones et ai. (Jones, G.L.,
Edmundson, H.M., Wesche, D. and Saul, A. (1991) Biochim. Biophys. Acta 1096, 33-40.) have now identified the
abnormality in the band 3 protein of ovalocytic red cell membranes. A major discovery in the Jones et al. study is the
presence of an extended peptide at the N-terminus of ovalocyte band 3 protein. This novel 13 amino acid extended
sequence is not found in the primary structure of normal band 3 protein and was suggested to be the cause of band 3
defect in ovalocytes. We have analyzed this extended sequence through Genbank using S W I S S - P R O T database and
found that an almost identical sequence exists in a malaria parasite protein called RESA.

Malaria in endemic areas of the tropics is a major
health problem with the rapid development of resis-
tance to antimalarials being a cause of great concern.
The occurrence of genetic red cell polymorphisms which
confer a certain degree of natural resistance to malarial
infection has been a subject of numerous studies. One
such polymorphism, called hereditary ovalocytosis, has
been observed in Papua New Guinea and parts of
Southeast Asia. Ovalocytic red cells received consider-
able attention when Kidson and his colleagues [1] first
reported dramatic resistance of these cells to P.
falciparum invasion. The resistance was later extended
to most of the species of malaria parasites which infect
primates [2].
Although it was shown that impaired cell deforma-
tion and increased ovalocyte membrane rigidity strongly
correlated with resistance to merozoite invasion, the
molecular basis of parasite invasion remained an enigma.
Recently, a very welcome development has been the
publication of two reports which begin to unravel the
mystery of the molecular abnormality in Melanesian
Correspondence: A.H. Chishti, Malaria Research Laboratory, Depart-
ment of Biomedical Research, St. Elizabeth's Hospital, Tufts Univer-
sity School of Medicine, Boston, MA 02135, U.S.A.
ovalocytosis [3,4]. Liu et al. [3] demonstrated that the
primary cause of the defect in ovalocytes lies in the
band 3 anion exchange protein which exhibited
abnormal proteinase cleavage. The structural defect is
dominantly inherited. The functional consequences of
the band 3 defect were traced to restricted lateral mobil-
ity and enhanced binding to ankyrin. Indeed an altered
interaction of band 3 with the underlying membrane
skeleton can explain increased rigidity and deformabil-
ity of the ovalocyte membrane. In retrospect, it should
be noted that the transmembrane band 3 protein has
been shown to be one of the erythrocyte receptors for P.
falciparum invasion in vitro [5]. Moreover, one cytoad-
hesive molecule on the surface of infected erythrocyte
has been shown to be structurally related to band 3
protein [6].
A more recent report by Jones et al. [4] identified an
altered N-terminus in the ovalocyte band 3 protein.
Markedly increased phosphorylation of the cytoplasmic
domain of band 3 was also observed. Again a structural
abnormality of band 3 protein was identified [4]. A
similar increased phosphorylation of ovalocyte band 3
was also recently noted by us (Chishti, Liu and Palek,
unpublished results). However what was surprising in
the Jones et al. [4] observations was the notion that the
slightly slower mobility of ovalocyte band 3 was due to
264
an extended N-terminus. The extension was unblocked
and a sequence of 13 amino acids, EERVEE-
PASDVQQ, was obtained by Edman degradation. The
N-terminus of normal band 3 is blocked and can not be
directly sequenced. The extended sequence is not found
in the known primary structure of normal band 3 pro-
tein. A suggestion was made that this new sequence may
have evolved by genetic duplication, since it resembles
the anionic character of the N-terminal sequence
( M E E L Q D D Y E D D ) of normal band 3 protein.
We analyzed the reported ovalocyte extended band 3
sequence by SWISS-PROT database (release number
16) and found it to be almost identical to a region of
ring-infected erythrocyte surface antigen (RESA) of the
human malaria parasite P. falciparum. The reported
peptide sequence is located at the 3' end of a repetitive
region in the middle of the RESA molecule [7]. The
sequence encoded in exon 2 of RESA from nucleotide
2492 to 2530 [7] is as follows:
RESA sequence: EEHVEEPASDVQQ
Ovalocyte band 3: EERVEEPASDVQQ
Our interpretations of this observation include: (a)
parasite contamination of ovalocyte specimens obtained
from malarial endemic areas. (b) An extremely tight
association of RESA with the cytoplasmic domain of
band 3. RESA is phosphorylated upon its interaction
with the erythrocyte membrane and may complicate
analysis of phosphorylated band 3 peptides. (c) A more
speculative scenario could still be an addilional ~c-
quence in ovalocyte band 3 mimicking epitope,~ ol
RESA. Indeed, such an argument can be supported b \
monoclonal antibodies which are known to crossreact
with both band 3 and RESA [8]. Moreover there is a
46.2% identity between a region (amino acids 28 41) of
the N-terminal domain of human band 3 and the RESA
sequence:
RESA sequence: EEHVEEPAS-DVQQ
H u m a n band 3: ESQMEEPAAHDTEA
Whatever turns out to be the explanation, the role of
band 3 protein in Melanesian ovalocytosis will surely be
a fascinating area of research in the near future.
References
1 Kidson, C., Lamont, G., Saul, A. and Nurse, G.T. (1981) Proc.
Natl. Acad. Sci. USA 78, 5829-5832.
2 Hadley, T., Saul, A., Lamont, G., Hudson, D.E., Miller, L.H. and
Kidson, C. (1983) J. Clin. Invest. 71, 780-782.
3 Liu, S-C., Zhai, S., Palek, J., Golan, D., Amato, D., Hassan, K.,
Nurse, G., Babona, D., Coetzer, T., Jarolim, P., Zaik, M. and
Borwein, S. (1990) N. Engl. J. Med. 323, 1530-1538.
4 Jones, G.L., Edmundson, H.M., Wesche, D. and Saul, A. (1991)
Biochim. Biophys. Acta 1096, 33-40.
5 0 k o y e , V.C.N. and Bennett, V. (1985) Science 227, 169- 171.
6 Winograd, E. and Sherman, I.W. (1989) J. Cell Biol. 108, 23-3(t.
7 Favaloro, J.M., Coppel, R.L., Corcoran, L.M., Foote, S.J., Brown,
G.V., Anders, R.F. and Kemp, D.J. (1986) Nucleic Acids Res. 14,
8265--8277.
8 Holmquist, G., Udomsangpetch, R., Berzins, K., Wigzell. H. and
Perlmann. P. (1988) Infect. lmmun. 56, 1545 1550.