Characterization and <i>N</i>-terminal sequence of human platelet proteoglycan

Périn, J.-P.; Bonnet, F; Maillet, Philippe; Jollès, Pierre

doi:10.1042/bj2551007

Cited by 38 publications

(12 citation statements)

References 15 publications

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“…Although the cDNA for serglycin predicts a molecular weight (MW) of 14.4 kd for the mature protein, it should be noted that the core protein from platelet serglycin and putative serglycins from hematopoietic tumor cells migrate at about twice this size on SDS-PAGE even in the presence of ␤-mercaptoethanol. 11,19,20 Part of the increase in apparent molecular weight is due to the approximately 4 to 6 residual hexasaccharide stubs, each 1.2 kd, expected to remain after the GAG depolymerizations, but the nonglycosylated core protein itself migrates at a higher apparent molecular weight than is expected from the …”

Section: Characterization Of 35 S-labeled Serglycin Synthesized By Humentioning

confidence: 99%

“…3 Over a number of years, it was found that several types of hematopoietic cells in several species synthesize a proteoglycan with a very small core protein and a characteristic resistance to trypsin digestion. [4][5][6][7][8][9][10] The proteoglycan core protein was purified from human platelets, 11 and its amino acid sequence and the complementary DNAs (cDNAs), which were cloned from human [12][13][14] and murine 15 hematopoietic cells, were found to be highly homologous to the rat L2 serglycin core protein. Messenger RNA (mRNA) for serglycin was subsequently identified in most blood and bone marrowderived cells by Northern blotting or in situ hybridization.…”

Section: Introductionmentioning

confidence: 99%

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Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells

Schick

Gradowski

Antonio

2001

Blood

102

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The serglycin proteoglycan is best known as a hematopoietic cell granule proteoglycan. It has been found that serglycin is synthesized by endothelial cells, is localized to cytoplasmic vesicles, and is constitutively secreted. Serglycin messenger RNA in human umbilical vein endothelial cells (HUVECs) and cultured human aortic endothelial cells was detected by reverse transcription-polymerase chain reaction. 35 Ssulfate-labeled secreted and intracellular proteoglycans were analyzed. It was found that 85% of the proteoglycans synthesized during culture were secreted. A core protein of the appropriate size for serglycin was detected by analysis of the chondroitinase-digested 35 Ssulfate-labeled HUVEC proteoglycans. This was the major core protein of the secreted chondroitin sulfate proteoglycans. Recombinant serglycin core protein was used to generate an antibody in chickens. A core protein identified by Western blotting of chondroitinase digests of HUVEC proteoglycans corresponded to the major 35 IntroductionThe serglycin proteoglycan was initially discovered as a secretory and membrane-associated product of rat L2 yolk sac tumor cells, 1 and its core protein was the first proteoglycan gene to be cloned. 2 L2 cells are thought to originate from parietal endoderm. 3 Over a number of years, it was found that several types of hematopoietic cells in several species synthesize a proteoglycan with a very small core protein and a characteristic resistance to trypsin digestion. [4][5][6][7][8][9][10] The proteoglycan core protein was purified from human platelets, 11 and its amino acid sequence and the complementary DNAs (cDNAs), which were cloned from human 12-14 and murine 15 hematopoietic cells, were found to be highly homologous to the rat L2 serglycin core protein. Messenger RNA (mRNA) for serglycin was subsequently identified in most blood and bone marrowderived cells by Northern blotting or in situ hybridization. 10,[16][17][18][19][20][21] Serglycin has thus come to be known as the hematopoietic proteoglycan. The serglycin proteoglycan is distinguished by the S/G (single-letter amino acid codes) repeat region in the central portion of the molecule, which is the site of attachment of the glycosaminoglycan (GAG) chains and gives the molecule its unique structural characteristics. The proteoglycan has either heparin or chondroitin sulfate GAGs, or it can be a hybrid of chondroitin sulfate and heparan sulfate chains, depending upon the cell source. [19][20][21] The function of the serglycin proteoglycan is not known, but it is likely that it is involved in packaging of proteins into secretory granules and/or directing the secretion of such molecules as cytokines or chymases. 22 These modes of regulation might reflect the interactions of the proteoglycan with other granule constituents or could involve osmotic effects that are due to the extensive hydration of the GAG chains. Serglycin is stored in granules for secretion upon activation by some cells and is secreted constitutively by others; in some cases, both mechan...

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Section: Characterization Of 35 S-labeled Serglycin Synthesized By Humentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells

Schick

Gradowski

Antonio

2001

Blood

102

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“…SG, cerebroglycan (with PRRLRL) (33), and perlecan (with TRRFRD) (34) are among the few PGs that contain heparin-binding consensus sequences on their core proteins. Interestingly, the SG core protein, which carries many heparin chains, migrates at twice its predicted molecular weight on PAGE gels under reducing conditions (35), suggesting dimerization. This could result from GAG chains of one PG binding to the core protein of another or from core-core associations through disulfide bonding.…”

Section: Figmentioning

confidence: 99%

Design of Peptides with High Affinities for Heparin and Endothelial Cell Proteoglycans

Verrecchio¹,

Germann²,

Schick³

et al. 2000

Journal of Biological Chemistry

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Proteoglycan-binding peptides were designed based on consensus sequences in heparin-binding proteins: XBBXBX and XBBBXXBX, where X and B are hydropathic and basic residues, respectively. Initial peptide constructs included (AKKARA) n and (ARKKAAKA) n (n ‫؍‬ 1-6). Affinity coelectrophoresis revealed that low M r peptides (600 -1300) had no affinities for low M r heparin, but higher M r peptides (2000 -3500) exhibited significant affinities (K d Х 50 -150 nM), which increased with peptide M r . Affinity was strongest when sequence arrays were contiguous and alanines and arginines occupied hydropathic and basic positions, but inclusion of prolines was disruptive. A peptide including a single consensus sequence of the serglycin proteoglycan core protein bound heparin strongly (K d Х 200 nM), likely owing to dimerization through cysteine-cysteine linkages. Circular dichroism showed that high affinity heparin-binding peptides converted from a charged coil to an ␣-helix upon heparin addition, whereas weak heparin-binding peptides did not. Higher M r peptides exhibited high affinities for total endothelial cell proteoglycans (K d Х 300 nM), and ϳ4-fold weaker affinities for their free glycosaminoglycan chains. Thus, peptides including concatamers of heparin-binding consensus sequences may exhibit strong affinities for heparin and proteoglycans. Such peptides may be applicable in promoting cell-substratum adhesion or in the design of drugs targeted to proteoglycan-containing cell surfaces and extracellular matrices. Proteoglycans (PGs)1 are composed of a core protein to which are covalently attached one or more sulfated glycosaminoglycans (GAGs). PGs are ubiquitous components of cell surfaces and the extracellular matrix, and their GAG chains contribute to myriad biological functions, such as modulation of enzyme activities, regulation of cell growth, and control of assembly of the extracellular matrix (1). PGs are thus potential targets for therapeutic intervention. For example, heparin antagonists are needed to take the place of protamine, a heterogeneous, sometimes toxic protein mixture commonly used to neutralize the anticoagulant activity of heparin in humans (2, 3); in the design of drugs to be targeted to PG-rich tissues, such as cartilage (4); and to be used to promote cell adhesion in a variety of situations, e.g. by promoting binding of cells that express abundant amounts of PGs, such as endothelial cells (5), to synthetic vascular graft surfaces. To develop a rationale for the design of such reagents, it is useful to examine known features of protein structure required for high affinity interactions with GAGs. Thus, analysis of the structural features of many known heparin-and heparan sulfate (HS)-binding proteins has revealed the presence of conserved motifs, through which GAG binding has been postulated to occur. Cardin and Weintraub (6) identified two clusters of basic charge in known heparin-binding proteins in which amino acids tend to be arranged in the patterns XBBXBX or XBBBXXBX, where B represents an ...

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“…SG mRNA expression and core protein have been detected in a multitude of cell types of hematopoietic origin, including mast cells, 9 macrophages, 10 T lymphocytes, 11 cytotoxic T lymphocytes (CTLs), 12,13 natural killer cells, 14 neutrophils, 15 and platelets. 8,16 In addition, SG is expressed by certain cells of nonhematopoietic origin (eg, parietal endoderm, 17,18 endothelial cells, 19 and murine uterine decidua 17 ).The name "serglycin" is derived from the long Ser-Gly repeat found in the central region of the core protein (ie, the GAG attachment region). The resultant dense clustering of the attached GAG chains is unique and likely explains the known protease resistance of SG.…”

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confidence: 99%

“…21 N-terminal sequencing of the platelet PG revealed its identity with SG. 16 Although it is reasonable to assume that platelet SG regulates storage of ␣-granule proteins, it has previously not been possible to determine the Submitted July 31, 2007; accepted December 14, 2007. Prepublished online as Blood First Edition paper, December 19, 2007; DOI 10.1182 DOI 10.…”

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confidence: 99%

Serglycin proteoglycan deletion induces defects in platelet aggregation and thrombus formation in mice

Woulfe

Lilliendahl

August

et al. 2008

Blood

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Serglycin (SG), the hematopoietic cell secretory granule proteoglycan, is crucial for storage of specific secretory proteins in mast cells, neutrophils, and cytotoxic T lymphocytes. We addressed the role of SG in platelets using SG-/- mice. Wild-type (WT) but not SG-/- platelets contained chondroitin sulfate proteoglycans. Electron microscopy revealed normal alpha-granule structure in SG-/- platelets. However, SG-/- platelets and megakaryocytes contained unusual scroll-like membranous inclusions, and SG-/- megakaryocytes showed extensive emperipolesis of neutrophils. SG-/- platelets had reduced ability to aggregate in response to low concentrations of collagen or PAR4 thrombin receptor agonist AYPGKF, and reduced fibrinogen binding after AYPGKF, but aggregated normally to ADP. 3H-serotonin and ATP secretion were greatly reduced in SG-/- platelets. The alpha-granule proteins platelet factor 4, beta-thromboglobulin, and platelet-derived growth factor were profoundly reduced in SG-/- platelets. Exposure of P-selectin and alphaIIb after thrombin treatment was similar in WT and SG-/- platelets. SG-/- mice exhibited reduced carotid artery thrombus formation after exposure to FeCl3. This study demonstrates that SG is crucial for platelet function and thrombus formation. We propose that SG-/- platelet function deficiencies are related to inadequate packaging and secretion of selected alpha-granule proteins and reduced secretion of dense granule contents critical for platelet activation.

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Characterization and N-terminal sequence of human platelet proteoglycan

Cited by 38 publications

References 15 publications

Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells

Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells

Design of Peptides with High Affinities for Heparin and Endothelial Cell Proteoglycans

Serglycin proteoglycan deletion induces defects in platelet aggregation and thrombus formation in mice

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