Vascular Biology |
From the Cell Biology of Diabetes Laboratory (P.J.L.), Baker Medical Research Institute, Melbourne, Victoria, Australia; the Division of Metabolism, Endocrinology and Nutrition (L.T., K.L.O., A.C.), Department of Medicine, and the Department of Pathology (T.N.W.), University of Washington, Seattle; and The Hope Heart Institute (T.N.W.), Seattle, Wash.
Correspondence to Thomas N. Wight, PhD, The Hope Heart Institute, 1124 Columbia Ave, Suite 783, Seattle, WA 98104. E-mail twight{at}hopeheart.org
| Abstract |
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Key Words: proteoglycans glycosaminoglycans smooth muscle cells transforming growth factor-ß1 lipoproteins
| Introduction |
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Transforming growth factor (TGF)-ß1 is a cytokine that is increased in atherogenesis and has been shown to promote atherosclerotic lesion formation.914 TGF-ß1 influences many of the events that contribute to lesion development. For example, TGF-ß1 increases the proliferation of arterial smooth muscle cells (ASMCs) under certain conditions11 and can affect cell differentiation and migration.15
In addition, TGF-ß1 affects the production of extracellular matrix components, such as proteoglycans, that contribute to atherosclerotic lesion progression.1618 For example, TGF-ß1 increases the synthesis by ASMCs of two proteoglycans, versican and biglycan,17,18 that accumulate in lesions of atherosclerosis.1923 In addition, these two proteoglycans have been localized to lipid-rich regions of atherosclerotic plaques along with TGF-ß1,13 suggesting a role for TGF-ß1 in lipid retention.
To determine whether TGF-ß1 affects proteoglycan-lipoprotein interactions and to identify the mechanism(s) responsible, we have assessed the capacity of proteoglycans produced by ASMCs in the presence or absence of TGF-ß1 to bind lipoproteins. We demonstrate that compared with proteoglycans isolated from ASMCs that were not treated with TGF-ß1, proteoglycans produced by TGF-ß1treated ASMCs exhibit enhanced binding to native human LDL. Furthermore, we present data to indicate that the effect of TGF-ß1 on proteoglycan-lipoprotein interactions is mainly through its effect of modulating the glycosaminoglycan synthetic machinery of the ASMC. The present study supports a proatherogenic role for TGF-ß1 through its ability to promote proteoglycan binding to LDL.
| Methods |
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Cell Culture
ASMCs isolated from monkey (Macaca nemestrina) thoracic aorta were kindly provided by the laboratory of Elaine Raines, University of Washington, Seattle, and cultured as previously described.24 Cells were grown in DMEM with 5.6 mmol/L glucose supplemented with pyruvate, nonessential amino acids, 4 mmol/L glutamine, antibiotics, and 5% calf serum. Cells were used between passages 6 and 18.
Determination of Proteoglycan and Xyloside-Initiated Glycosaminoglycan Synthesis
Cells were seeded at 60 000 cells per well into 12-well cell culture dishes in DMEM (5.6 mmol/L glucose) containing 10% serum. The medium was changed on the following day, and cells were confluent by day 3 or 4. Cells were made quiescent by changing to DMEM with 0.1% serum and incubating for 48 hours. Cells were radiolabeled with 50 µCi/mL [35S]sulfate (to label glycosaminoglycan chains) or 60 µCi/mL [35S]methionine (to label core proteins) in the presence or absence of TGF-ß1 (2 ng/mL) for 24 hours. Earlier studies showed that this concentration provided maximal stimulation of proteoglycan synthesis by monkey ASMCs.17,18 To determine whether TGF-ß1 influenced glycosaminoglycan synthesis independent of core protein synthesis, TGF-ß1treated cultures were supplemented with 0.5 mmol/L xyloside during the 24-hour labeling period.25 Xyloside acts as an artificial acceptor for galactosyltransferase in the initiation of glycosaminoglycan synthesis and results in increased synthesis and secretion of free glycosaminoglycan chains and decreased synthesis of intact proteoglycans.25 At the conclusion of the incubation period, culture medium (50 µL) from each well was spotted in duplicate onto paper (Whatman 3M, Whatman Int), which was then washed (5 times) in cetylpyridinium chloride (CPC) solution (1% in 0.05 mol/L NaCl). The paper was air-dried, cut into segments, and counted in a scintillation vial with Instagel (Packard Corp) for determination of CPC-precipitable counts as a measure of glycosaminoglycan synthesis.26
Proteoglycan Isolation and Characterization
Proteoglycans from triplicate wells of 12-well plates were passed over DEAE-Sephacel (0.25 mL) and washed with "low salt buffer" (0.25 mol/L NaCl, 8 mol/L urea, 2 mmol/L Na2EDTA, and 0.5% Triton X-100, pH 7.5). Proteoglycans were eluted with "high salt buffer" (3 mol/L NaCl, 8 mol/L urea, 2 mmol/L Na2EDTA, and 0.5% Triton X-100, pH 7.5) 5 times at 0.3 mL. Aliquots (10 µL) were counted to locate the fractions containing proteoglycans, which were then combined. Aliquots of the pools were precipitated with ethanol by using chondroitin sulfate as a carrier and finally dissolved for SDS-PAGE in 20 µL of buffer (8 mol/L urea and 2 mmol/L Na2EDTA, pH 7.0).27
Pooled proteoglycans eluted from DEAE-Sephacel minicolumns were treated with 1 mol/L sodium borohydride in 50 mmol/L NaOH for 4 hours at 45°C to release glycosaminoglycan chains from core proteins by reductive ß elimination.17,18 The reaction was terminated by neutralization with glacial acetic acid. The glycosaminoglycans obtained from untreated and TGF-ß1treated cells were prepared for SDS-PAGE, hydrodynamic sizing, and gel mobility shift assays. Proteoglycans and glycosaminoglycans were subjected to electrophoresis (SDS-PAGE) by using a 4% to 12% gradient acrylamide gel for proteoglycans and a 4% to 16% gradient acrylamide gel for glycosaminoglycans with a 3.5% stacking gel according to the procedure of Laemmli.28 Apparent molecular weights were estimated by comparisons to 14C protein standards (GIBCO-BRL Life Technologies Inc).
Hydrodynamic sizes of the DEAE-purified proteoglycans and glycosaminoglycans were determined by molecular sieve chromatography on analytical Sepharose CL-2B or CL-6B columns (0.7x110 cm), respectively, equilibrated in 8 mol/L urea buffer with 0.5% Triton X-100.17,18,27 Eluted fractions (1 mL) were collected, and radioactivity was determined by liquid scintillation counting. The elution position of the free radioisotope was used as a marker for the total volume, and the void volume was determined by the elution position of [3H]DNA. Assessment of glycosaminoglycan molecular weight was made by chromatography on Sepharose CL-6B columns by using the standard curve of Wasteson.29
Proteoglycan-Lipoprotein Binding Assay
LDLs (density 1.019 to 1.063 g/mL) were separated from plasma from normal human volunteers by preparative ultracentrifugation in a Beckman VTi 50 vertical rotor (Beckman Instruments) and purified by sequential density gradient ultracentrifugation.30 LDL was stored in the dark at 4°C in 1 mol/L EDTA until it was used in the gel mobility shift assay.
Radiolabeled proteoglycans were isolated as described above and concentrated by using Centricon concentrators (Millipore Corp). Cellulose filtration membranes with cutoffs of 50 and 10 kDa were used for proteoglycans and free glycosaminoglycans, respectively. The sample was diluted with HEPES buffer (10 mmol/L HEPES, 140 mmol/L NaCl, 5 mmol/L CaCl2, and 2 mmol/L MgCl2, pH 7.4) and recentrifuged. The concentration and dilution process was repeated 4 times.
The interaction of proteoglycans with lipoproteins was assessed by using a gel mobility shift assay.31 Briefly, fixed amounts of radiolabeled proteoglycans, glycosaminoglycans, or core proteins were incubated with increasing concentrations of LDL for 1 hour at 37°C in the HEPES buffer. The reaction mixture was subjected to electrophoresis at 60 V for 3 hours at 4°C in 0.7% NuSieve Agarose Gel (FMC Bioproducts) in circulating running buffer (10 mmol/L HEPES, 2 mmol/L CaCl2, and 4 mmol/L MgCl2, pH 7.2). The gels were fixed with CPC (0.1% [wt/vol] in 70% ethanol) for 1 hour and air-dried. Radioactivity was assessed by using PhosphorImager (Molecular Dynamics) screens. The images were analyzed by using Optiquant proprietary software. The amount of complexed versus unbound proteoglycans or glycosaminoglycans in each lane was assessed, and the percent bound was calculated as the proportion of the radioactivity retained at the origin of the gel relative to the total radioactivity in each lane. Values for Kd and Bmax from the electrophoretic data were determined with the SAAM II modeling program (SAAM Institute) by using the Michaelis-Menten equation.
To determine whether core proteins of proteoglycans had bound to LDL, [35S]methionine-labeled proteoglycans were isolated as described above. Samples were dialyzed into 0.1 mol/L Tris HCl, 10 mmol/L calcium acetate, and 18 mmol/L sodium acetate, pH 7.0. Glycosaminoglycan chains were digested from aliquots of control and TGF-ßlstimulated proteoglycans by digestion with 0.7 U/mL chondroitin ABC lyase (ICN), 20 U/mL heparinase I, and 20 U/mL heparinase II in the presence of protease inhibitors for 6 hours at 37°C. Additional aliquots were treated in parallel in the absence of enzymes. The digestion was dialyzed into gel mobility shift buffer by using MOPS (20 mmol/L MOPS, 140 mmol/L NaCl, 5 mmol/L CaCl2, and 2 mmol/L MgCl2, pH 7.4), and binding to lipoproteins was determined by a gel mobility shift assay as described above. SDS-PAGE confirmed that the digestion of glycosaminoglycan chains was complete (data not shown).
| Results |
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Analysis of the gels demonstrates that proteoglycan-lipoprotein complexes form at lower concentrations of LDL with proteoglycans from the TGF-ß1treated ASMCs compared with proteoglycans from control ASMCs (Figure 1A and 1B). Michaelis-Menten analysis of the binding curves fitted the data with correlation coefficiets >0.95. The total binding was nearly identical in control and TGF-ß1treated preparations and approximated 100% (Figure 1C). The Kd value for binding of the control proteoglycan pool was 81 µg/mL of LDL (Kd 1.47x10-7 mol/L apoB), assuming a molecular mass of 550 000 Da for apoB, and the Kd for binding of proteoglycans from the TGF-ß1treated cells was
18 µg/mL LDL (3.27x10-8 mol/L apoB, P<0.002). The lower Kd value indicates higher affinity binding of the proteoglycans synthesized by ASMCs treated with TGF-ß1.
To determine whether the glycosaminoglycan chains were responsible for binding to LDL, the glycosaminoglycan chains were chemically released from the core proteins of the proteoglycans, and their binding to LDL was investigated. Earlier studies showed that the addition of TGF-ßl to ASMCs caused a lengthening of the glycosaminoglycan chains on the chondroitin sulfate and dermatan sulfate proteoglycans synthesized by ASMCs,17,18 and these differences were confirmed in the present study (data not shown). The glycosaminoglycan chains released from proteoglycans were bound to LDL in a concentration-dependent manner (Figure 2). The binding affinity for glycosaminoglycan chains released from ASMC proteoglycans treated with TGF-ßl was higher for LDL compared with that for glycosaminoglycan chains from untreated cells (Figure 2, P=0.04).
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Because previous and present data have suggested that TGF-ß1mediated elongation of glycosaminoglycan chains and increased binding affinity may be related, further experiments were undertaken to determine whether the effect of TGF-ß1 was due to the effects on glycosaminoglycan chain elongation. By use of ß-D-xyloside (an initiator of glycosaminoglycan synthesis), it is possible to examine glycosaminoglycan synthesis in the absence of core protein synthesis. Thus, xyloside stimulates glycosaminoglycan synthesis but reduces proteoglycan synthesis in ASMCs.25 ß-D-Xyloside caused an
2-fold increase in the incorporation of radiolabeled sulfate into glycosaminoglycans during a 24-hour labeling period compared with control cells (Figure 3). TGF-ß1 treatment enhanced this increase, suggesting that TGF-ß1 can have an effect on the glycosaminoglycan synthetic machinery in the absence of proteoglycan core protein synthesis.
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To further evaluate the xylosideTGF-ß1 effect, newly synthesized proteoglycans and glycosaminoglycans were evaluated by SDS-PAGE and molecular sieve chromatography (Figure 4). In the controls, xyloside caused a reduction in the amount of proteoglycans that separate at the top of the SDS-polyacrylamide gel and the appearance of a broad band of radioactivity between 18 and 43 kDa (Figure 4A). Previous studies have identified the high molecular weight band as versican and the lower molecular weight broad band as a mixture of chondroitin sulfate and dermatan sulfate chains.17,18,25 In the TGF-ß1treated cultures, xyloside also reduced high molecular weight proteoglycans but shifted the broad band of radioactivity to a range of 29 to 80 kDa, suggesting an increase in the molecular weight of components found in this band (Figure 4A). To further assess the effect of TGF-ß1 on the xyloside-initiated glycosaminoglycan synthesis, the preparations were hydrodynamically sized by using Sepharose CL-6B columns. Three peaks of radioactivity were observed in the media of control and TGF-ß1treated ASMCs. The control ASMC cultures contained peaks at Kav 0.00 (peak I), Kav 0.25 (peak II, shoulder), and Kav 0.51 (peak III) (Figure 4). Similar peaks of radioactivity were observed for the preparations from TGF-ß1treated ASMCs, with the exception that peak III was shifted to an earlier elution position, indicating a larger sized population of molecules (Kav 0.43). Our previous studies have shown that peaks I and II represent intact proteoglycans and that peak III contains a mixture of chondroitin sulfate and dermatan sulfate chains.17,18,25 The shift in peak III caused by TGF-ß1 treatment corresponds to an increase in average molecular mass from 19.2 to 29.0 kDa.29 These data confirm that TGF-ß1 can increase the length of glycosaminoglycan chains in the absence of core protein synthesis.
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To determine whether the longer xyloside-initiated glycosaminoglycan chains produced in the presence of TGF-ß1 were bound more avidly to LDL, the gel mobility shift assay was performed by using radiolabeled samples from the xyloside-treated preparations. The xyloside-initiated chains are smaller than glycosaminoglycan chains on intact proteoglycans in ASMCs (data not shown).25 Xyloside-initiated chains from control cultures were bound to LDL with a Kd value of 1350 µg/mL LDL (2.45x10-6 mol/L apoB), whereas xyloside-initiated chains isolated from cultures treated with TGF-ß1 were bound to LDL with a Kd value of 880 µg/mL LDL (1.6x10-6 mol/L apoB, P=0.03), again indicating that TGF-ßl results in the synthesis of glycosaminoglycan chains with increased binding affinity for LDL (Figure 5).
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Gel mobility shift assays were also performed on control and TGF-ß1stimulated proteoglycans labeled with [35S]methionine, which had been treated with chondroitin ABC lyase and heparinase I and II to remove the glycosaminoglycans chains. No binding was seen between LDL and core proteins synthesized under control conditions or in the presence of TGF-ß1 (data now shown).
| Discussion |
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Elucidating the mechanism by which TGF-ß1 causes an increase in glycosaminoglycan chain length is a potential therapeutic target for intervention in the pathological process of binding and retention of lipoproteins within the vascular wall. These studies support a proatherogenic role for TGF-ß1.
| Acknowledgments |
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Received October 4, 2001; accepted October 8, 2001.
| References |
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2.
Braunwald E. Shattuck lecture: cardiovascular medicine at the turn of the millennium: triumphs, concerns, and opportunities. N Engl J Med. 1997; 337: 13601369.
3. Wei M, Haffner SM, Gaskill SP, Stern MP. Effects of diabetes and level of glycemia on all-cause and cardiovascular mortality. Diabetes Care. 1998; 21: 11671172.[Abstract]
4.
Radhakrishnamurthy B, Srinivasan SR, Vijayagopal P, Berenson GS. Arterial wall proteoglycans: biological properties related to pathogenesis of atherosclerosis. Eur Heart J. 1990; 11 (suppl E): 148157.
5.
Williams KJ, Tabas I. The response to retention hypothesis of early atherogenesis. Arterioscler Thromb Vasc Biol. 1995; 15: 551561.
6.
Hurt-Camejo E, Olsson U, Wiklund O, Bondjers G, Camejo G. Cellular consequences of the association of apoB lipoproteins with proteoglycans: potential contribution to atherogenesis. Arterioscler Thromb Vasc Biol. 1997; 17: 10111017.
7. Camejo G, Hurt-Camejo E, Wiklund O, Bondjers G. Association of apo B lipoproteins with arterial proteoglycans: pathological significance and molecular basis. Atherosclerosis. 1998; 139: 205222.[CrossRef][Medline] [Order article via Infotrieve]
8. Chait A, Wight TN. Interaction of native and modified low-density lipoproteins with extracellular matrix. Curr Opin Lipidol. 2000; 11: 457463.[CrossRef][Medline] [Order article via Infotrieve]
9. Majesky MW, Lindner V, Twardzik DR, Schwartz SM, Reidy MA. Production of transforming growth factor beta 1 during repair of arterial injury. J Clin Invest. 1991; 88: 904910.[Medline] [Order article via Infotrieve]
10.
Nabel EG, Shum L, Ponpili VJ, Yang ZY, San H, Shu HB, Liptay S, Gold L, Gordon D, Derynck R, et al. Direct gene transfer of transforming growth factor ß1 gene into arteries stimulates fibrocellular hyperplasia. Proc Natl Acad Sci U S A. 1993; 90: 1075910763.
11.
Agrotis A, Saltis J, Bobik A. Transforming growth factor-beta 1 gene activation and growth of smooth muscle from hypertensive rats. Hypertension. 1994; 23: 593599.
12.
Bobik A, Agrotis A, Kanellakis P, Dilley R, Krushinsky A, Smirnov V, Tararak E, Condron M, Kostolias G. Distinct patterns of transforming growth factor-beta isoform and receptor expression in human atherosclerotic lesions: colocalization implicates TGF-beta in fibrofatty lesion development. Circulation. 1999; 99: 28832891.
13. Evanko SP, Raines EW, Ross R, Gold LI, Wight TN. Proteoglycan distribution in lesions of atherosclerosis depends on lesion severity, structural characteristics, and the proximity of platelet-derived growth factor and transforming growth factor-beta. Am J Pathol. 1998; 152: 533546.[Abstract]
14. McCaffrey TA. TGF-betas and TGF-beta receptors in atherosclerosis. Cytokine Growth Factor Rev. 2000; 11: 103114.[CrossRef][Medline] [Order article via Infotrieve]
15.
Christen T, Bochaton-Piallat ML, Neuville P, Rensen S, Redard M, van Eys G, Gabbiani G. Cultured porcine coronary artery smooth muscle cells: a new model with advanced differentiation. Circ Res. 1999; 85: 99107.
16. Scott L, Kerr A, Haydock D, Merrilees M. Subendothelial proteoglycan synthesis and transforming growth factor beta distribution correlate with susceptibility to atherosclerosis. J Vasc Res. 1997; 34: 365377.[Medline] [Order article via Infotrieve]
17.
Schönherr E, Järveläinen HT, Sandell LJ, Wight TN. Effects of platelet-derived growth factor and transforming growth factor-beta 1 on the synthesis of a large versican-like chondroitin sulfate proteoglycan by arterial smooth muscle cells. J Biol Chem. 1991; 266: 1764017647.
18.
Schönherr E, Järveläinen HT, Kinsella MG, Sandell LJ, Wight TN. Platelet-derived growth factor and transforming growth factor-beta 1 differentially affect the synthesis of biglycan and decorin by monkey arterial smooth muscle cells. Arterioscler Thromb. 1993; 13: 10261036.
19. Lin H, Ignatescu M, Wilson JE, Roberts CR, Horley KJ, Winters GL, Costanzo MR, McManus BM. Prominence of apolipoproteins B, (a) and E in the intimae of coronary arteries in transplanted human hearts: geographic relationship to vessel wall proteoglycans. J Heart Lung Transplant. 1996; 15: 12231232.[Medline] [Order article via Infotrieve]
20.
OBrien KD, Olin KL, Alpers CE, Chiu W, Ferguson M, Hudkins K, Wight TN, Chait A. Comparison of apolipoprotein and proteoglycan deposits in human coronary atherosclerotic plaques: colocalization of biglycan with apolipoproteins. Circulation. 1998; 98: 519527.
21. Gutierrez P, OBrien KD, Ferguson M, Nikkari ST, Alpers CE, Wight TN. Differences in the distribution of versican, decorin, and biglycan in atherosclerotic human coronary arteries. Cardiovasc Pathol. 1997; 6: 271278.[CrossRef]
22.
Halpert I, Sires U, Potter-Perigo S, Wight TN, Shapiro DS, Welgus HG, Wickline SA, Parks WC. Matrilysin is expressed by lipid-laden macrophages at sites of potential rupture in atherosclerotic lesions and localized to areas of versican deposits. Proc Natl Acad Sci U S A. 1996; 93: 97489753.
23. Riessen R, Isner JM, Blessing E, Loushin D, Nikol S, Wight TN. Regional differences in the distribution of the proteoglycans, biglycan, and decorin in the extracellular matrix of atherosclerotic and restenotic human coronary arteries. Am J Pathol. 1994; 144: 962974.[Abstract]
24.
Stout RW, Bierman EL, Ross R. Effect of insulin on the proliferation of cultured primate arterial smooth muscle cells. Circ Res. 1975; 36: 319327.
25. Potter-Perigo S, Braun KR, Schönherr E, Wight TN. Altered proteoglycan synthesis via the false acceptor pathway can be dissociated from beta-D-xyloside inhibition of proliferation. Arch Biochem Biophys. 1992; 297: 101109.[CrossRef][Medline] [Order article via Infotrieve]
26. Wasteson A, Uthne K, Westermark B. A novel assay for the biosynthesis of sulfated polysaccharide and its application to studies on the effects of somatomedin on cultured cells. Biochem J. 1973; 136: 10691074.[Medline] [Order article via Infotrieve]
27. Yanagishita M, Midura RJ, Hascall VC. Proteoglycans: isolation and purification from tissue cultures. Methods Enzymol. 1987; 138: 279289.[Medline] [Order article via Infotrieve]
28. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227: 680685.[CrossRef][Medline] [Order article via Infotrieve]
29. Wasteson A. A method for the determination of the molecular weight and molecular-weight distribution of chondroitin sulfate. J Chromatogr. 1971; 59: 8797.[CrossRef][Medline] [Order article via Infotrieve]
30. Heinecke JW, Baker L, Rosen H, Chait A. Superoxide-mediated modification of low density lipoprotein by arterial smooth muscle cells. J Clin Invest. 1986; 77: 757761.[Medline] [Order article via Infotrieve]
31. Hurt-Camejo E, Camejo G, Sartipy P. Measurements of proteoglycan-lipoprotein interaction by gel mobility shift assay. Methods Mol Biol. 1998; 110: 267279.[Medline] [Order article via Infotrieve]
32.
Cardoso LE, Mourao PA. Glycosaminoglycan fractions from human arteries presenting diverse susceptibilities to atherosclerosis have different binding affinities to plasma LDL. Arterioscler Thromb. 1994; 14: 115124.
33.
Chang MY, Potter-Perigo S, Tsoi C, Chait A, Wight TN. Oxidized low density lipoproteins regulate synthesis of monkey aortic smooth muscle cell proteoglycans that have enhanced native low density lipoprotein binding properties. J Biol Chem. 2000; 275: 47664773.
34.
Camejo G, Fager G, Rosengren B, Hurt-Camejo E, Bondjers G. Binding of low density lipoproteins by proteoglycans synthesized by proliferating and quiescent human arterial smooth muscle cells. J Biol Chem. 1993; 268: 1413114137.
35. Srinivasan SR, Xy JH, Vijayagopal P, Radhakrishnamurthy B, Berenson GS. Injury to the arterial wall of rabbits produces proteoglycan variants with enhanced low-density lipoprotein-binding property. Biochim Biophys Acta. 1993; 1168: 158166.[Medline] [Order article via Infotrieve]
36. Westergren-Thorsson G, Schmidtchen A, Sarnstrand B, Fransson LA, Malmström A. Transforming growth factor-beta induces selective increase of proteoglycan production and changes in the copolymeric structure of dermatan sulphate in human skin fibroblasts. Eur J Biochem. 1992; 205: 277286.[Medline] [Order article via Infotrieve]
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