|
|
 Previous Article | Table of Contents | Next Article 
Biologic and biochemic properties of recombinant platelet factor 4
demonstrate identity with the native protein
KS Park, S Rifat, H Eck, K Adachi, S Surrey and M Poncz
Division of Hematology, Children's Hospital of Philadelphia, PA 19104.
Platelet factor 4 (PF4) is a 70 amino acid protein released from the
alpha-granules of platelets after activation. The exact biologic function
of this protein is unknown. We have constructed an expression vector for
recombinant PF4 (rPF4) in the T7-based promoter vector pT7-7 to better
study the relationship between PF4 structure and function. The protein was
expressed in Escherichia coli and purified to homogeneity by
heparin-agarose affinity chromatography and reverse- phase high-performance
liquid chromatography. Purity of protein was confirmed by immunoblot
analysis and sodium dodecyl sulfate- polyacrylamide gel electrophoresis,
which resulted in a single component with a molecular weight of 8,000
daltons. The amino acid composition and sequence of the N-terminal 20
residues showed that rPF4 is identical to PF4 prepared from human platelets
(hPF4), except for an N-terminal initiating methionine residue. Immunoblots
revealed that rPF4 and hPF4 bound polyclonal anti-hPF4 equally well, while
chemotaxis experiments demonstrated similar potencies as neutrophil
attractants. Dose-dependent neutrophil chemotactic responses and
competitive studies with polyclonal anti-hPF4 antiserum further demonstrate
similar chemotactic properties of the two PF4 species. In conclusion, our
data show that this recombinant protein and the native protein appears to
have similar immunologic, heparin-binding, and chemotactic properties. The
chemotactic properties of hPF4 appear to be entirely intrinsic to the
protein and not due, in part, to any contaminating protein. Furthermore,
our expression vector should prove useful for the construction of
recombinant forms of PF4 to investigate structure/function relationships of
this biologically important protein.
Volume 75,
Issue 6,
pp. 1290-1295,
03/15/1990
Copyright © 1990 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
M. P. Lambert, L. Rauova, M. Bailey, M. C. Sola-Visner, M. A. Kowalska, and M. Poncz
Platelet factor 4 is a negative autocrine in vivo regulator of megakaryopoiesis: clinical and therapeutic implications
Blood,
August 15, 2007;
110(4):
1153 - 1160.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Rauova, L. Zhai, M. A. Kowalska, G. M. Arepally, D. B. Cines, and M. Poncz
Role of platelet surface PF4 antigenic complexes in heparin-induced thrombocytopenia pathogenesis: diagnostic and therapeutic implications
Blood,
March 15, 2006;
107(6):
2346 - 2353.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Rauova, M. Poncz, S. E. McKenzie, M. P. Reilly, G. Arepally, J. W. Weisel, C. Nagaswami, D. B. Cines, and B. S. Sachais
Ultralarge complexes of PF4 and heparin are central to the pathogenesis of heparin-induced thrombocytopenia
Blood,
January 1, 2005;
105(1):
131 - 138.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. E. Eslin, C. Zhang, K. J. Samuels, L. Rauova, L. Zhai, S. Niewiarowski, D. B. Cines, M. Poncz, and M. A. Kowalska
Transgenic mice studies demonstrate a role for platelet factor 4 in thrombosis: dissociation between anticoagulant and antithrombotic effect of heparin
Blood,
November 15, 2004;
104(10):
3173 - 3180.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. S. Sachais, A. Kuo, T. Nassar, J. Morgan, K. Kariko, K. J. Williams, M. Feldman, M. Aviram, N. Shah, L. Jarett, et al.
Platelet factor 4 binds to low-density lipoprotein receptors and disrupts the endocytic itinerary, resulting in retention of low-density lipoprotein on the cell surface
Blood,
May 15, 2002;
99(10):
3613 - 3622.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. Q. Li, W. Liu, K. S. Park, B. S. Sachais, G. M. Arepally, D. B. Cines, and M. Poncz
Defining a second epitope for heparin-induced thrombocytopenia/thrombosis antibodies using KKO, a murine HIT-like monoclonal antibody
Blood,
February 15, 2002;
99(4):
1230 - 1236.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. M. Arepally, S. Kamei, K. S. Park, K. Kamei, Z. Q. Li, W. Liu, D. L. Siegel, W. Kisiel, D. B. Cines, and M. Poncz
Characterization of a murine monoclonal antibody that mimics heparin-induced thrombocytopenia antibodies
Blood,
March 1, 2000;
95(5):
1533 - 1540.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Ziporen, Z.Q. Li, K.S. Park, P. Sabnekar, W.Y. Liu, G. Arepally, Y. Shoenfeld, T. Kieber-Emmons, D.B. Cines, and M. Poncz
Defining an Antigenic Epitope on Platelet Factor 4 Associated With Heparin-Induced Thrombocytopenia
Blood,
November 1, 1998;
92(9):
3250 - 3259.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. A.-R. Higazi, E. Lavi, K. Bdeir, A. M. Ulrich, D. G. Jamieson, D. J. Rader, D. C. Usher, W. Kane, T. Ganz, and D. B. Cines
Defensin Stimulates the Binding of Lipoprotein (a) to Human Vascular Endothelial and Smooth Muscle Cells
Blood,
June 15, 1997;
89(12):
4290 - 4298.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. L. Bauer, G. Arepally, B. A. Konkle, B. Mestichelli, S. S. Shapiro, D. B. Cines, M. Poncz, S. McNulty, J. Amiral, W. W. Hauck, et al.
Prevalence of Heparin-Associated Antibodies Without Thrombosis in Patients Undergoing Cardiopulmonary Bypass Surgery
Circulation,
March 4, 1997;
95(5):
1242 - 1246.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
G. Arepally, S. E. McKenzie, X.-M. Jiang, M. Poncz, and D. B. Cines
Fcgamma RIIA H/R131 Polymorphism, Subclass-Specific IgG Anti-Heparin/Platelet Factor 4 Antibodies and Clinical Course in Patients With Heparin-Induced Thrombocytopenia and Thrombosis
Blood,
January 15, 1997;
89(2):
370 - 375.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. G. Malkowski, J. Y. Wu, P. H. Johnson, and B. F. P. Edwards
The Crystal Structure of Recombinant Human Neutrophil-activating Peptide-2 (M6L) at 1.9-Ĺ Resolution
J. Biol. Chem.,
March 31, 1995;
270(13):
7077 - 7087.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
