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Volume 272, Number 52, Issue of December 26, 1997
pp. 32817-32823
(Received for publication, August 6, 1997, and in revised form, October 6, 1997)
PTX3 is a prototypic long pentraxin consisting of
a C-terminal 203-amino acid pentraxin-like domain coupled with an
N-terminal 178-amino acid unrelated portion. The present study was
designed to characterize the structure and ligand binding properties of human PTX3, in comparison with the classical pentraxins C-reactive protein and serum amyloid P component. Sequencing of Chinese hamster ovary cell-expressed PTX3 revealed that the mature secreted protein starts at residue 18 (Glu). Lectin binding and treatment with N-glycosidase F showed that PTX3 is
N-glycosylated, sugars accounting for 5 kDa of the monomer
mass (45 kDa). Circular dichroism analysis indicated that the protein
consists predominantly of
Multimer Formation and Ligand Recognition by the Long
Pentraxin PTX3
SIMILARITIES AND DIFFERENCES WITH THE SHORT PENTRAXINS
C-REACTIVE PROTEIN AND SERUM AMYLOID P COMPONENT
,
,
,
,
,
,
,
,
,
,
,
and§§
From the Istituto di Ricerche Farmacologiche "Mario
Negri," Via Eritrea 62, 20157 Milano, Italy; ¶ Istituto di
Ostetricia e Ginecologia, IRCCS Burlo Garofolo, Trieste, Italy;
DOMPE' S.P.A., L'Aquila, Italy; ** Istituto di Scienze
Farmacologiche, Università di Milano, Milan, Italy;
Dipartimento di Fisiologia e Patologia,
Università di Trieste, Trieste, Italy; and
§§ Sezione di Patologia e Immunologia,
Dipartimento di Biotecnologie, Università di Brescia,
Brescia, Italy
-sheets with a minor 
-helical
component. While in gel filtration the protein is eluted with a
molecular mass of 
900 kDa, gel electrophoresis using nondenaturing,
nonreducing conditions revealed that PTX3 forms multimers predominantly
of 440 kDa apparent molecular mass, corresponding to decamers, and that
disulfide bonds are required for multimer formation. The ligand binding
properties of PTX3 were then examined. As predicted based on modeling,
inductive coupled plasma/atomic emission spectroscopy showed that PTX3
does not have coordinated Ca2+. Unlike the classical
pentraxins CRP and SAP, PTX3 did not bind phosphoethanolamine,
phosphocholine, or high pyruvate agarose. PTX3 in solution, bound to
immobilized C1q, but not C1s, and, reciprocally, C1q bound to
immobilized PTX3. Binding of PTX3 to C1q is specific and saturable with
a Kd 7.4 × 10
8 M as
determined by solid phase binding assay. The Chinese hamster ovary
cell-expressed pentraxin domain bound C1q when multimerized. Thus, as
predicted on the basis of computer modeling, the prototypic long
pentraxin PTX3 forms multimers, which differ from those formed by
classical pentraxins in terms of protomer composition and requirement for disulfide bonds, and does not recognize CRP/SAP ligands. The capacity to bind C1q, mediated by the pentraxin domain, is consistent with the view that PTX3, produced in tissues by endothelial cells or
macrophages in response to interleukin-1 and tumor necrosis factor, may
act as a local regulator of innate immunity.
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D. G. Souza, A. C. Soares, V. Pinho, H. Torloni, L. F. L. Reis, M. T. Martins, and A. A. M. Dias Increased Mortality and Inflammation in Tumor Necrosis Factor-Stimulated Gene-14 Transgenic Mice after Ischemia and Reperfusion Injury Am. J. Pathol., May 1, 2002; 160(5): 1755 - 1765. [Abstract] [Full Text] [PDF]
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A. A. M. Dias, A. R. Goodman, J. L. Dos Santos, R. N. Gomes, A. Altmeyer, P. T. Bozza, M. de Fatima Horta, J. Vilcek, and L. F. L. Reis TSG-14 transgenic mice have improved survival to endotoxemia and to CLP-induced sepsis J. Leukoc. Biol., June 1, 2001; 69(6): 928 - 936. [Abstract] [Full Text] [PDF]
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P. Rovere, G. Peri, F. Fazzini, B. Bottazzi, A. Doni, A. Bondanza, V. S. Zimmermann, C. Garlanda, U. Fascio, M. G. Sabbadini, et al. The long pentraxin PTX3 binds to apoptotic cells and regulates their clearance by antigen-presenting dendritic cells Blood, December 15, 2000; 96(13): 4300 - 4306. [Abstract] [Full Text] [PDF]
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G. Peri, M. Introna, D. Corradi, G. Iacuitti, S. Signorini, F. Avanzini, F. Pizzetti, A. P. Maggioni, T. Moccetti, M. Metra, et al. PTX3, A Prototypical Long Pentraxin, Is an Early Indicator of Acute Myocardial Infarction in Humans Circulation, August 8, 2000; 102(6): 636 - 641. [Abstract] [Full Text] [PDF]
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T. M. Misenheimer, K. G. Huwiler, D. S. Annis, and D. F. Mosher Physical Characterization of the Procollagen Module of Human Thrombospondin 1 Expressed in Insect Cells J. Biol. Chem., December 22, 2000; 275(52): 40938 - 40945. [Abstract] [Full Text] [PDF]
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