Biochem. J. (1992) 287, 291-297
Characterization of the human properdin gene
(Printed in Great Britain)
Kathleen F. NOLAN, Stefan KALUZ,* Jonathan M. G. HIGGINS, Dimitrios GOUNDIS
and Kenneth B. M. REID
MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OXI 3QU, U.K.
A cosmid clone containing the complete coding sequence of the human properdin gene has been characterized. The gene
is located at one end ofthe
Two discrepancies with the published cDNA sequence [Nolan, Schwaeble, Kaluz, Dierich & Reid (1991) Eur. J. Immunol.
21, 771-776] have been resolved. Properdin has previously been described as a modular protein, with the majority of its
sequence composed of six tandem repeats of a sequence motif of
sequence (TSR), initially described in thrombospondin [Lawler & Hynes (1986) J. Cell Biol. 103, 1635-1648; Goundis
& Reid (1988), Nature (London) 335, 82-85]. Analysis of the genomic sequence data indicates that the human properdin
gene is organized into ten exons which span
(phase 1-1), which supports the hypothesis that modular proteins evolved by a process involving exon shuffling. TSR1 is
also coded for by a discrete exon, but the boundaries are asymmetrical (phase 2-1). The sequence coding for the sixth TSR
is split across the final two exons of the gene with the first 38 amino acids of the repeat coded for by an asymmetric exon
(phase 1-2). This split at the genomic level has been shown, by alignment analysis, to be reflected at the protein level with
the division of repeat 6 into TSR-like and TSR-unlike sequences.
- 40 kb cosmid insert and
- 8.2 kb ofthe sequence data have been obtained from this region.
- 60 amino acids which is related to the type-I repeat
- 6 kb ofthe genome. TSRs 2-5 are coded for by discrete, symmetrical exons
Properdin is a basic glycoprotein which is present in the serum,
at a concentration of
mainly dimers, trimers and tetramers, in the proportions
26: 54:20 (Pangburn, 1989). These polymericforms arecomposed
1981). In electron micrographs the composite monomers appear
as flexible rods (each
interact through their N- and C-terminal regions in a head-to-tail
manner (Smith et al., 1984). The polymerization is thought to be
an early, intracellular event andmonomers have notbeendetected
in serum (Farries & Atkinson, 1989). Although the interactions
between monomers are non-covalent (Minta & Lepow, 1974),
they are strong, as illustrated by the stability of the polymeric
forms when isolated individually (Pangburn, 1989).
Properdin was first described by Pillemer et al. (1954) and is
the only known positive regulator of the alternative pathway of
complement. In contrast to the classical pathway ofcomplement
activation, which is mainly antibody-dependent, the alternative
pathway is activated by a wide range of surfaces in an antibody-
independent manner. This pathway forms an amplification loop
which leads to the deposition oflarge numbers ofC3b molecules
on activating surfaces. TheseC3b molecules function as opsonins,
promoting recognition and uptake by phagocytic cells, and also
act as foci for the formation of cytolytic membrane attack
complexes by the terminal complement components. Properdin
promotes C3b-dependent functions by binding to and stabilizing
the inherently labile C3/C5 convertase complexes, C3bBb and
C3bnBb,present on activating surfaces. This binding promotes
the cleavage and activation of components C3 and C5 and also
inhibits binding of the negative regulatory enzyme factor I
(Farries et al., 1988).
Deficiency of functional properdin is inherited as an X-linked,
recessive disorder and the symptoms exhibited by patients with
- 5 mg/l, as a mixture of cyclic polymers,
- 53 kDa, asymmetric monomers (Reid & Gagnon,
- 26 nm long) and are considered to
such a deficiency (reviewed by Sjoholm, 1990) illustrate the
importance of properdin in host defence against bacterial,
particularly neisserial, infections. The properdin gene has been
localized to the short arm of the X chromosome, in the region
Xpll.23-Xpll.3 (Goundis et al., 1989; Coleman et al., 1991).
Genetic linkage studies have also mapped the properdin-
deficiency locus to this region (Goonewardena et al., 1988),
suggesting that this condition results from genetic lesion(s)
located within, or very close to, the properdin structural gene.
Previous analysis of mouse and human cDNA sequences
(Goundis & Reid, 1988; Nolan et al., 1991) indicated that, like
many of the other complement components, properdin is a
modular protein (Reid & Day, 1989). The human monomer is
442 amino acids long and possesses a single, potential N-linked
glycosylation site at residue 401 (Nolan et al., 1991). Analysis of
the derived amino acid sequences of both mouse and human
properdin indicated the presence of distinct N- (49 amino acid)
and C- (29 amino acid) terminal regions flanking a central
repetitive region composed of six tandemly repeated sequence
repeat sequence (TSR),
(Lawler & Hynes, 1986). TSR modules have also been reported
in the terminal complement components C6, C7, C8a, C8,8and
C9 (see references in DiScipio & Hugli, 1989), human and
hamster homologues of thrombospondin (TS) (Paul et al., 1989;
thrombospondin-related anonymous protein (TRAP) protein of
Plasmodium falciparum (Robson et al., 1988) and the SSP2
protein ofPlasmodium yoelii (Hedstrom et al., 1990). A region of
sequence similarity with the N-terminal region of the repeat has
also been reported in the circumsporozoite protein of a number
ofmalaria parasites (Ozaki et al., 1983; Dame et al., 1984; Arnot
et al., 1985; Eichinger et al., 1986; Galinski et al., 1987; Lal et al.,
1987, 1988). Although these proteins have diverse biological
functions and the role of the TSR in any of these proteins is not
- 60 amino acids long and related to the type-I
first identified in thrombospondin
Abbreviations used: TSR, type-I repeat sequence; TS, thrombospondin; TRAP, thrombospondin-related anonymous protein; SSC, 0.15M-
NaCl/0.015 M-sodium citrate; PMA, phorbol 12-myristate 13-acetate; ORF, open reading frame; RACE, rapid amplificationofcDNA ends.
* Present address: Institute of Virology, 84246 Bratislava, Czechoslovakia.
Human properdin gene
reported to induce rearrangements such as deletions, as in the
case of the low-density-lipoprotein receptor and Cl inhibitor
genes (Lehrman et al., 1987; Tosi et al., 1989), gross deletions of
the properdin gene in deficient patients have not been detected
(Nolan et al., 1989). In the case of the X-linked dysfunctional
form of the protein a defect in the structural properdin gene is
directly implied. The recently reported polymorphic dinucleotide
repeat sequence, identified
perdin gene in Cos4XP (Coleman et al., 1991), provides a means
for detecting individuals within a family who have inherited the
defective gene (Kolble et al., 1991) and, in combination with this
method of detection, the present report of the properdin gene
sequence should be of considerable
investigating the underlying genetic defects responsible for the
different forms of properdin deficiency.
- 15 kb downstream from the pro-
This work was funded by the Medical Research Council, U.K.
J.M.G.H. is supported by a Wellcome Trust Prize Studentship. We
thank Beryl Moffat for tissue culture work, Dr. A. J. Day for advice on
the alignment analyses and Dr. R. D. Campbell for critical reading of
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Received 29 October 1991/24 February 1992; accepted 19 March 1992