Crystal Structure of Recombinant Human Platelet Factor 4

Zhang, Xiaohua; Chen, Liqing; Bancroft, Daniel P.; Lai, Ching-san; Maione, Theodore E.

doi:10.1021/bi00193a025

Cited by 185 publications

(204 citation statements)

References 31 publications

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“…In solution structures of macrophage inflammatory protein-1␤ (28) and RANTES (29,30), which are CC chemokines, N-terminal residues constitute the dimer interface. The crystal structure of platelet factor-4, a CXC chemokine, showed it to be a tetramer, and it has two dimer interfaces similar to NAP-2, an A/B interface like IL-8 and an A/C interface like RANTES (31,32). By necessity, the crystals of NAP-2 were grown at high concentrations of the protein (17 mg/ml in 100 mM sodium acetate, 200 mM ammonium acetate, pH 4.6), and this could be the reason for the formation of the tetramer.…”

Section: Resultsmentioning

confidence: 99%

Neutrophil-activating Peptide-2 and Melanoma Growth-stimulatory Activity Are Functional as Monomers for Neutrophil Activation

Rajarathnam

Kay

Dewald

et al. 1997

Journal of Biological Chemistry

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Neutrophil-activating peptide-2 (NAP-2) and melanoma growth-stimulatory activity (MGSA) are members of the chemokine family of inflammatory proteins. The structures of NAP-2, determined by x-ray crystallography, and MGSA, elucidated by NMR spectroscopy, revealed a tetramer and dimer, respectively. In order to address the relevance of multimeric species to their activities on neutrophils, analogs of NAP-2 and MGSA were synthesized in which the backbone amide proton of Leu-22 in NAP-2, and Val-26 in MGSA, was substituted with the bulky methyl group (NH 3 NCH 3 ). These analogs were shown to be monomeric by sedimentation equilibrium ultracentrifugation studies and were similar to the corresponding native protein in assays for neutrophil elastase release and Ca 2؉ mobilization from IL-8R1 and IL-8R2 transformed cells. Sedimentation equilibrium studies of the native NAP-2 and MGSA were also carried out to address the association behavior. For NAP-2, there was no evidence for the tetramer, but an equilibrium between monomers and dimers and the dissociation constant was calculated to be 50 -100 M. Similarly, MGSA showed a monomer-dimer equilibrium with a K d of ϳ5 M. The data from the monomeric analogs and also the calculation of dissociation constants indicate that NAP-2 and MGSA have a tendency to associate above the concentrations required for maximal activity or for receptor activation, but at functional concentrations they are predominantly monomers.Recruitment and accumulation of circulating leukocytes at the site of inflammation are mediated in part by a family of small molecular weight proteins called chemokines (1-3). They have four conserved cysteines and are called either CXC or CC chemokines, depending on whether the first two cysteines are separated by one amino acid (CXC) or are adjacent (CC). Functionally, the CXC chemokines predominantly attract neutrophils, whereas CC chemokines attract monocytes, lymphocytes, and eosinophils.Three well studied proteins of the CXC chemokine family are interleukin-8 (IL-8) 1

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Section: Resultsmentioning

confidence: 99%

Neutrophil-activating Peptide-2 and Melanoma Growth-stimulatory Activity Are Functional as Monomers for Neutrophil Activation

Rajarathnam

Kay

Dewald

et al. 1997

Journal of Biological Chemistry

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“…The PF4 ZIP peptide, a GCN4-based coiled-coil motif designed to mimic the lysine-rich heparinbinding region of PF4 [34,58], was employed in these studies, with ultimate goals of expanding the number and types of binding interactions employed in hydrogel assembly and to manipulate materials properties and delivery profiles. The use of PF4 ZIP was motivated by the fact that platelet factor 4 (PF4) is perhaps the strongest heparin-binding protein known [59]. The fact that the PF4 ZIP peptide also binds strongly to heparin was confirmed via affinity chromatography and SPR experiments.…”

Section: Discussionmentioning

confidence: 99%

Manipulation of hydrogel assembly and growth factor delivery via the use of peptide–polysaccharide interactions

Zhang

Furst

Kiick

2006

Journal of Controlled Release

117

131

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The design of materials in which assembly, mechanical response, and biological properties are controlled by protein-polysaccharide interactions could provide materials that mimic the biological environment and find use in the delivery of growth factors. In the investigations reported here, a heparin-binding, coiled-coil peptide, PF4 ZIP , was employed to mediate the assembly of heparinized polymers. The heparin-binding affinity of this peptide was compared with that of other heparinbinding peptides (HBP) via heparin-sepharose chromatography and surface plasmon resonance (SPR) experiments. Results from these experiments indicate that PF4 ZIP demonstrates a higher heparin-binding affinity and heparin association rate when compared to the heparin-binding domains of antithrombin III (ATIII) and heparin-interacting protein (HIP). Viscoelastic hydrogels were formed upon the association of PF4 ZIP -functionalized star poly(ethylene glycol) (PEG-PF4 ZIP ) with low-molecular-weight heparin-functionalized star PEG (PEG-LMWH). The viscoelastic properties of the hydrogels can be altered via variations in the ratio of LMWH to PF4 ZIP . bFGF release from these gels have also been investigated. Comparison of the bFGF release profiles with the hydrogel erosion profiles indicates that bFGF delivery from this class of hydrogels is mainly an erosioncontrolled process and the rates of bFGF release can be modulated via choice of HBP or via variations in the mechanical properties of the hydrogels. Manipulation of hydrogel physical properties and erosion profiles will provide novel materials for controlled growth factor delivery and other biomedical applications.

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“…fragments, the 5Ј sequences of which correspond to the C termini of the PF4 deleted fragments PF4 1-60 , PF4 , and PF4 1-40 , PF4 , PF4 [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] , PF4 [1][2][3][4][5][6][7][8][9][10][11][12][13] , PF4 ⌬41-50 , and PF4 ⌬45-48 (Fig. 1B).…”

Section: Methodsmentioning

confidence: 99%

“…PF4 is an antiangiogenic factor that inhibits megakaryocyte and endothelial cell proliferation (15,16). Importantly, the PF4 structure has been solved by x-ray cristallography, allowing structure-function predictions (17,18). PF4 is released along with other ␣-granule proteins upon physiological and pathological stimulation (9).…”

mentioning

confidence: 99%

Evidence for a Granule Targeting Sequence within Platelet Factor 4

Issertial

Rosa²,

Briquet-Laugier³

2005

Journal of Biological Chemistry

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Platelets achieve bleeding arrest at sites of vascular injury via secretion of secretory proteins from their storage granules, termed ␣-granules. We have recently analyzed granule targeting of platelet factor 4 (PF4), a secretory ␣-granule chemokine, and demonstrated that PF4 ␣-granule storage relied upon determinants within PF4 mature sequence. To define these determinants, PF4 mutants fused to the fluorescent reporter protein green fluorescent protein were generated by progressive deletions and site-directed mutagenesis. They were then transfected in AtT20 cells and assessed for granule targeting by colocalization with ACTH-containing granules, using laser scanning confocal microscopy. This strategy identified the amino acid 41-50 (LIATLKNGRK) sequence as most critical for PF4 granule targeting and/or storage; its deletion from PF4 induced a marked decrease in granule storage (from 81 ؎ 2% to 17 ؎ 3%, p < 0.0001). Ala-scanning mutagenesis of LIATLKNGRK narrowed down the targeting motif to LKNG. A direct role for LKNG in ␣-granule targeting was confirmed in the thrombopoietin-induced human megakaryocytic Dami cells, in which the LKNGgreen fluorescent protein chimera exhibited an 82.5 ؎ 1.8% colocalization with the ␣-granule proteins von Willebrand factor and P-selectin. LKNG is poorly conserved within the chemokine family. However three-dimensional alignments of the human ␣-granule chemokines Nap-2 (neutrophil-activating peptide) and RANTES (Regulated upon Activation Normal T Cell Expressed and Secreted) with PF4 revealed that LKNG, a surface-exposed hydrophilic turn/loop, matched Nap-2 (LKDG) and RANTES (TRKN) peptides with similar features. Moreover Nap-2 and RANTES peptides exhibited the same ␣-granule targeting efficiency than LKNG. We therefore postulate that the three-dimensional and physicochemical characteristics of PF4 LKNG are of general relevance to ␣-granule targeting of chemokines and possibly of other ␣-granule proteins.

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Crystal Structure of Recombinant Human Platelet Factor 4

Cited by 185 publications

References 31 publications

Neutrophil-activating Peptide-2 and Melanoma Growth-stimulatory Activity Are Functional as Monomers for Neutrophil Activation

Neutrophil-activating Peptide-2 and Melanoma Growth-stimulatory Activity Are Functional as Monomers for Neutrophil Activation

Manipulation of hydrogel assembly and growth factor delivery via the use of peptide–polysaccharide interactions

Evidence for a Granule Targeting Sequence within Platelet Factor 4

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