MCP-1 binds to oxidized LDL and is carried by lipoprotein(a) in human plasma

Lipoprotein oxidation plays an important role in pathogenesis of atherosclerosis. Oxidized low density lipoprotein (OxLDL) induces profound inflammatory responses in vascular cells, such as production of monocyte chemoattractant protein-1 (MCP-1) [chemokine (C-C motif) ligand 2], a key chemokine in the initiation and progression of vascular inflammation. Here we demonstrate that OxLDL also binds MCP-1 and that the OxLDL-bound MCP-1 retains its ability to recruit monocytes. A human MCP-1 mutant in which basic amino acids Arg-18 and Lys-19 were replaced with Ala did not bind to OxLDL. The MCP-1 binding to OxLDL was inhibited by the monoclonal antibody E06, which binds oxidized phospholipids (OxPLs) in OxLDL. Because OxPLs are carried by lipoprotein(a) [Lp(a)] in human plasma, we tested to determine whether Lp(a) binds MCP-1. Recombinant wild-type but not mutant MCP-1 added to human plasma bound to Lp(a), and its binding was inhibited by E06. Lp(a) captured from human plasma contained MCP-1 and the Lp(a)-associated endogenous MCP-1 induced monocyte migration. These results demonstrate that OxLDL and Lp(a) bind MCP-1 in vitro and in vivo and that OxPLs are major determinants of the MCP-1 binding. The association of MCP-1 with OxLDL and Lp(a) may play a role in modulating monocyte trafficking during atherogenesis. Lipoprotein oxidation plays an important role in pathogenesis of atherosclerosis. Oxidized low density lipoprotein (OxLDL) induces profound inflammatory responses in vascular cells, such as production of monocyte chemoattractant protein-1 (MCP-1) [chemokine (C-C motif) ligand 2], a key chemokine in the initiation and progression of vascular inflammation. Here we demonstrate that OxLDL also binds MCP-1 and that the OxLDL-bound MCP-1 retains its ability to recruit monocytes. A human MCP-1 mutant in which basic amino acids Arg-18 and Lys-19 were replaced with Ala did not bind to OxLDL. The MCP-1 binding to OxLDL was inhibited by the monoclonal antibody E06, which binds oxidized phospholipids (OxPLs) in OxLDL. Because OxPLs are carried by lipoprotein(a) [Lp(a)] in human plasma, we tested to determine whether Lp(a) binds MCP-1. Recombinant wild-type but not mutant MCP-1 added to human plasma bound to Lp(a), and its binding was inhibited by E06. Lp(a) captured from human plasma contained MCP-1 and the Lp(a)-associated endogenous MCP-1 induced monocyte migration. These results demonstrate that OxLDL and Lp(a) bind MCP-1 in vitro and in vivo and that OxPLs are major determinants of the MCP-1 binding. The association of MCP-1 with OxLDL and Lp(a) may play a role in modulating monocyte trafficking during atherogenesis. Vascular cells secrete chemokines into the extravascular space. Glycosaminoglycans (GAGs) are expressed on the surface of endothelial cells, where they bind and transcytose chemokines to the luminal surface (1Salanga C.L. Handel T.M. Chemokine oligomerization and interactions with receptors and glycosaminoglycans: the role of structural dynamics in function.Exp. Cell Res. 2011; 317: 590-601Crossref PubMed Scopus (132) Google Scholar, 2Weber C. Noels H. Atherosclerosis: current pathogenesis and therapeutic options.Nat. Med. 2011; 17: 1410-1422Crossref PubMed Scopus (1559) Google Scholar). Monocyte chemoattractant protein-1 (MCP-1) [synonym: chemokine (C-C motif) ligand 2 (CCL2)], is a major chemokine involved in development of atherosclerosis via monocyte recruitment to the vascular wall (3Boring L. Gosling J. Cleary M. Charo I.F. Decreased lesion formation in CCR2-/- mice reveals a role for chemokines in the initiation of atherosclerosis.Nature. 1998; 394: 894-897Crossref PubMed Scopus (1680) Google Scholar). Plasma levels of MCP-1 are associated with traditional risk factors for atherosclerosis in the general population and with an increased risk for death or myocardial infarction (MI) in patients with acute coronary syndrome (4de Lemos J.A. Morrow D.A. Sabatine M.S. Murphy S.A. Gibson C.M. Antman E.M. McCabe C.H. Cannon C.P. Braunwald E. Association between plasma levels of monocyte chemoattractant protein-1 and long-term clinical outcomes in patients with acute coronary syndromes.Circulation. 2003; 107: 690-695Crossref PubMed Scopus (391) Google Scholar, 5Deo R. Khera A. McGuire D.K. Murphy S.A. Meo Neto J.P. Morrow D.A. de Lemos J.A. Association among plasma levels of monocyte chemoattractant protein-1, traditional cardiovascular risk factors, and subclinical atherosclerosis.J. Am. Coll. Cardiol. 2004; 44: 1812-1818Crossref PubMed Scopus (239) Google Scholar, 6de Lemos J.A. Morrow D.A. Blazing M.A. Jarolim P. Wiviott S.D. Sabatine M.S. Califf R.M. Braunwald E. Serial measurement of monocyte chemoattractant protein-1 after acute coronary syndromes: results from the A to Z trial.J. Am. Coll. Cardiol. 2007; 50: 2117-2124Crossref PubMed Scopus (133) Google Scholar). GAGs have been shown to play an important role in the in vivo activation and function of MCP-1 (7Proudfoot A.E. Handel T.M. Johnson Z. Lau E.K. LiWang P. Clark-Lewis I. Borlat F. Wells T.N. Kosco-Vilbois M.H. Glycosaminoglycan binding and oligomerization are essential for the in vivo activity of certain chemokines.Proc. Natl. Acad. Sci. USA. 2003; 100: 1885-1890Crossref PubMed Scopus (643) Google Scholar, 8Weber K.S. von Hundelshausen P. Clark-Lewis I. Weber P.C. Weber C. Differential immobilization and hierarchical involvement of chemokines in monocyte arrest and transmigration on inflamed endothelium in shear flow.Eur. J. Immunol. 1999; 29: 700-712Crossref PubMed Scopus (182) Google Scholar). Earlier studies demonstrated that negatively charged GAGs bind to MCP-1 via the basic amino acids Arg-18 and Lys-19 in the MCP-1 molecule (9Lau E.K. Paavola C.D. Johnson Z. Gaudry J.P. Geretti E. Borlat F. Kungl A.J. Proudfoot A.E. Handel T.M. Identification of the glycosaminoglycan binding site of the CC chemokine, MCP-1: implications for structure and function in vivo.J. Biol. Chem. 2004; 279: 22294-22305Abstract Full Text Full Text PDF PubMed Scopus (170) Google Scholar). Oxidized low density lipoprotein (OxLDL) is an electronegative component of vascular lesions and an important pathogenic factor in the development of atherosclerosis (10Glass C.K. Witztum J.L. Atherosclerosis. The road ahead.Cell. 2001; 104: 503-516Abstract Full Text Full Text PDF PubMed Scopus (2636) Google Scholar). OxLDL activates vascular cells to secrete MCP-1 (11Reddy S. Hama S. Grijalva V. Hassan K. Mottahedeh R. Hough G. Wadleigh D.J. Navab M. Fogelman A.M. Mitogen-activated protein kinase phosphatase 1 activity is necessary for oxidized phospholipids to induce monocyte chemotactic activity in human aortic endothelial cells.J. Biol. Chem. 2001; 276: 17030-17035Abstract Full Text Full Text PDF PubMed Scopus (47) Google Scholar), leading to recruitment of monocytes, which differentiate into macrophages and internalize OxLDL. The resulting lipid-laden macrophage foam cells are a hallmark of atherosclerotic lesions that play a central role in atherosclerosis progression. We hypothesized that, similar to MCP-1 binding to GAGs, MCP-1 would also bind to electronegative OxLDL, which in turn would play a role in guiding monocyte recruitment. Native LDL (nLDL) (density = 1.019–1.063 g/ml) was isolated from plasma of normolipidemic donors by sequential ultracentrifugation (12Havel R.J. Bragdon J.H. Eder H.A. The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum.J. Clin. Invest. 1955; 34: 1345-1353Crossref PubMed Scopus (6485) Google Scholar). Contamination of native and modified LDL preparations by endotoxin was assessed with a LAL QCL-1000 kit (Lonza). LDL preparations with LPS higher than 50 pg/mg protein were discarded. To produce OxLDL, 0.1 mg/ml of nLDL was incubated with 10 μM CuSO4 for 18 h at 37°C (13Miller Y.I. Viriyakosol S. Binder C.J. Feramisco J.R. Kirkland T.N. Witztum J.L. Minimally modified LDL binds to CD14, induces macrophage spreading via TLR4/MD-2, and inhibits phagocytosis of apoptotic cells.J. Biol. Chem. 2003; 278: 1561-1568Abstract Full Text Full Text PDF PubMed Scopus (336) Google Scholar). The extent of LDL oxidation was assessed by measuring thiobarbituric acid-reactive substances (typically, more than 30 nmol/mg protein), and OxLDL was concentrated to 1 mg/ml using a 100 kDa cut off centrifugal concentrator (Millipore) and sterile filtered (0.22 μm). Plasma samples (n = 127) were collected from patients presenting with chest pain and suspected acute coronary syndromes (ST-segment elevation MI; non-ST-segment elevation MI and unstable angina) on admission to the Veteran's Affairs Medical Center San Diego. Patients that ultimately ruled out for MI by clinical criteria and myocardial enzyme biomarkers were included as controls. The blood was immediately spun down in EDTA and the plasma separated and stored at −70°C. The collection of human plasma and the assays on these samples were approved by the Veteran's Affairs Medical Center and the University of California, San Diego Human Research Subjects Protection Programs, respectively, and all participants gave written informed consent. C57BL6/J mice were wild type or transgenic expressing human apoB-100, human apo(a), or lipoprotein(a) [Lp(a)], i.e., both apoB-100 and apo(a), as previously reported (14Schneider M. Witztum J.L. Young S.G. Ludwig E.H. Miller E.R. Tsimikas S. Curtiss L.K. Marcovina S.M. Taylor J.M. Lawn R.M. et al.High-level lipoprotein [a] expression in transgenic mice: evidence for oxidized phospholipids in lipoprotein [a] but not in low density lipoproteins.J. Lipid Res. 2005; 46: 769-778Abstract Full Text Full Text PDF PubMed Scopus (51) Google Scholar, 15Merki E. Graham M.J. Mullick A.E. Miller E.R. Crooke R.M. Pitas R.E. Witztum J.L. Tsimikas S. Antisense oligonucleotide directed to human apolipoprotein B-100 reduces lipoprotein(a) levels and oxidized phospholipids on human apolipoprotein B-100 particles in lipoprotein(a) transgenic mice.Circulation. 2008; 118: 743-753Crossref PubMed Scopus (129) Google Scholar, 16Merki E. Graham M. Taleb A. Leibundgut G. Yang X. Miller E.R. Fu W. Mullick A.E. Lee R. Willeit P. et al.Antisense oligonucleotide lowers plasma levels of apolipoprotein (a) and lipoprotein (a) in transgenic mice.J. Am. Coll. Cardiol. 2011; 57: 1611-1621Crossref PubMed Scopus (105) Google Scholar). Mice were housed in a barrier facility with a 12 h light/12 h dark cycle, and fed normal mouse chow containing 4.5% fat (Harlan Teklad). All animal experiments were approved by the University of California, San Diego Institutional Animal Care and Use Committee. Wild-type and R18A/K19A mutant MCP-1 constructs were expressed in Escherichia coli and purified by reverse-phase HPLC as previously described (9Lau E.K. Paavola C.D. Johnson Z. Gaudry J.P. Geretti E. Borlat F. Kungl A.J. Proudfoot A.E. Handel T.M. Identification of the glycosaminoglycan binding site of the CC chemokine, MCP-1: implications for structure and function in vivo.J. Biol. Chem. 2004; 279: 22294-22305Abstract Full Text Full Text PDF PubMed Scopus (170) Google Scholar, 17Paavola C.D. Hemmerich S. Grunberger D. I. A. R. M. S. D. L. et monocyte chemoattractant protein-1 (MCP-1) binds and activates the MCP-1 Biol. Chem. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar). The MCP-1 preparations were tested for endotoxin with a LAL QCL-1000 kit (Lonza). were in all MCP-1 nLDL and OxLDL samples were incubated with MCP-1 for 30 at 37°C they were on a and at using an of were collected and for MCP-1 and apoB-100 using as described of OxLDL, with wild-type mutant S. D.A. Miller E. H. G. J.A. P. Curtiss L.K. et for oxidized phospholipids or oxidized macrophage of oxidized lipoproteins.J. Clin. Invest. 1999; PubMed Scopus Google were on a with for 18 h at 100 was in the or the The were to a the was with in and incubated with an antibody or an antibody monoclonal antibody S.G. Witztum J.L. Curtiss L.K. S. of the low density lipoprotein of by a monoclonal PubMed Google for human The was and incubated with a antibody directed the incubated with for and with an Lp(a) binding were with antibody S. Lau Miller E.R. A. Curtiss L.K. Witztum J.L. coronary results in acute in oxidized phospholipids and and long-term responses to oxidized 2004; PubMed Scopus Google at were and with for Plasma samples for human or for mouse were in and incubated for at were and incubated with 50 antibody and for at were incubated with for at and incubated with in for at The were with an and results were as 100 or oxidized were on as described S. D.A. Miller E. H. G. J.A. P. Curtiss L.K. et for oxidized phospholipids or oxidized macrophage of oxidized lipoproteins.J. Clin. Invest. 1999; PubMed Scopus Google and MCP-1 binding was as was from and was by oxidation of as previously described Navab M. S. Witztum J.L. W. D. W. et by of oxidized phospholipids in oxidized low density lipoprotein that induce interactions and evidence for in vivo.J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). A transmigration was with human cells using and the MCP-1 nLDL or OxLDL in 10 were in the and 1 cells were added to the experiments cells were with 100 a chemokine type 2 R.J. R. D.J. Yang G. S. H. et of as a of CC chemokine 2 Med. Chem. 2008; PubMed Scopus Google Scholar, S. M. S. M. P. R. of on human 2011; PubMed Scopus Google Scholar), for 30 the of the and the was in the for the of the The cells were to for 2 h at and were in and with were in of the were in and to a of Lp(a) was isolated from human plasma using the antibody S. Lau Miller E.R. A. Curtiss L.K. Witztum J.L. coronary results in acute in oxidized phospholipids and and long-term responses to oxidized 2004; PubMed Scopus Google on plasma samples were with of MCP-1 the Lp(a) were added to human plasma and incubated for 2 h at to endogenous plasma in was incubated with 2 of the monoclonal antibody S. Lau Miller E.R. A. Curtiss L.K. Witztum J.L. coronary results in acute in oxidized phospholipids and and long-term responses to oxidized 2004; PubMed Scopus Google at with by a 1 h with 50 of at The were and in the as a chemoattractant in the To in they were 10 for the of the Because of in the the of cells MCP-1 was in of basic amino acids in binding of MCP-1 to OxPLs in OxLDL. OxLDL was with or 10 of wild-type or mutant MCP-1 for 30 at 37°C and the samples were on native and with to apoB-100 or MCP-1. samples were with of or MCP-1 binding to of MCP-1 to or was tested in a are as (n = binding to Plasma samples from wild-type or transgenic mice expressing human apoB-100, human apo(a), or both were incubated with MCP-1 for 30 at and to with a antibody and a MCP-1 A human plasma with an Lp(a) of was and incubated with wild-type or mutant MCP-1 for 30 at and to as in are as of The was with plasma samples from with similar A human plasma with an Lp(a) of was and incubated with or and with wild-type MCP-1 for 30 at The samples were to as in are as of The was with plasma samples from with similar A human plasma with an Lp(a) of was and incubated with of wild-type MCP-1 for 30 at in the of or 100 The samples were to as in are as of The was with plasma samples from with similar The as in was to endogenous levels of in plasma samples from cardiovascular to 2 plasma samples in the of Lp(a) from plasma samples in was on with an the were as a chemoattractant in a The of in plasma samples A and were and A of plasma was with MCP-1 to the Lp(a) cells were with 100 for 30 the of the are as of The in between the MCP-1 samples in is to the as described in was at and assays were in and the results are as of assays were by and results of MCP-1 binding were by with with were To the that MCP-1 binds to OxLDL, MCP-1 was with OxLDL or a and were for apoB-100 and MCP-1 shown in MCP-1 bound OxLDL to a with MCP-1 associated with OxLDL also its to induce of and the of cells in to OxLDL-bound MCP-1 was higher than in to MCP-1. The of monocyte was in experiments with an of the MCP-1 MCP-1 binding to OxLDL was in a native in which MCP-1 with the to wild-type an MCP-1 mutant in which the basic amino acids Arg-18 and Lys-19 were replaced with did not bind to OxLDL. the E06, which binds and oxidized phospholipids (OxPLs) on the surface of OxLDL S. D.A. Miller E. H. G. J.A. P. Curtiss L.K. et for oxidized phospholipids or oxidized macrophage of oxidized lipoproteins.J. Clin. Invest. 1999; PubMed Scopus Google Scholar), binding of wild-type MCP-1 to OxLDL. To demonstrate that MCP-1 binds to we added MCP-1 to in which or were shown in MCP-1 bound to a of Navab M. S. Witztum J.L. W. D. W. et by of oxidized phospholipids in oxidized low density lipoprotein that induce interactions and evidence for in vivo.J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), but not to OxLDL in vivo in the is that such OxLDL in the we have shown that OxPLs are in plasma and that Lp(a) is the lipoprotein that a major of OxPLs in human plasma C. A. A. E. Miller E.R. M.J. Binder C.J. S. R.M. M.J. et function of lipoprotein [a] as a of oxidized phospholipids in human Lipid Res. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, S. J.L. as a risk factor of cardiovascular a for increased to its and Am. Coll. Cardiol. PubMed Scopus Google Scholar). Because the antibody MCP-1 binding to OxLDL and MCP-1 bound to OxPLs we tested to determine whether Lp(a) binds MCP-1 as we tested plasma samples from transgenic mice expressing human apoB-100, human apo(a), or Lp(a) both apoB-100 and using a with an and an MCP-1 was previously shown that the human and mouse particles of these mice of OxPLs and that of the OxPLs in plasma are carried by (14Schneider M. Witztum J.L. Young S.G. Ludwig E.H. Miller E.R. Tsimikas S. Curtiss L.K. Marcovina S.M. Taylor J.M. Lawn R.M. et al.High-level lipoprotein [a] expression in transgenic mice: evidence for oxidized phospholipids in lipoprotein [a] but not in low density lipoproteins.J. Lipid Res. 2005; 46: 769-778Abstract Full Text Full Text PDF PubMed Scopus (51) Google Scholar, 15Merki E. Graham M.J. Mullick A.E. Miller E.R. Crooke R.M. Pitas R.E. Witztum J.L. Tsimikas S. Antisense oligonucleotide directed to human apolipoprotein B-100 reduces lipoprotein(a) levels and oxidized phospholipids on human apolipoprotein B-100 particles in lipoprotein(a) transgenic mice.Circulation. 2008; 118: 743-753Crossref PubMed Scopus (129) Google Scholar, 16Merki E. Graham M. Taleb A. Leibundgut G. Yang X. Miller E.R. Fu W. Mullick A.E. Lee R. Willeit P. et al.Antisense oligonucleotide lowers plasma levels of apolipoprotein (a) and lipoprotein (a) in transgenic mice.J. Am. Coll. Cardiol. 2011; 57: 1611-1621Crossref PubMed Scopus (105) Google Scholar), similar to A. Witztum J.L. Tsimikas S. Oxidized phospholipids on a cardiovascular and cardiovascular Med. 2011; PubMed Scopus (133) Google Scholar). was endogenous MCP-1 on Lp(a), which was that the mice were after MCP-1 to mouse plasma we MCP-1 on lipoproteins the with human plasma we also binding of wild-type but not mutant R18A/K19A to Lp(a) of inhibited MCP-1 binding to Lp(a) in the human plasma Because MCP-1 binds to GAGs (1Salanga C.L. Handel T.M. Chemokine oligomerization and interactions with receptors and glycosaminoglycans: the role of structural dynamics in function.Exp. Cell Res. 2011; 317: 590-601Crossref PubMed Scopus (132) Google Scholar), we tested to determine whether MCP-1 binding to Lp(a) by a to human plasma inhibited MCP-1 binding to Lp(a) in a that GAGs and Lp(a) may for an MCP-1 and determine of the MCP-1 associated with Lp(a) was in a of patients presenting with chest using the in which we captured Lp(a) from human plasma and for the of MCP-1. The of in samples as as from to in of the We to determine whether the Lp(a)-associated endogenous MCP-1 induced monocyte migration. Lp(a) was from plasma with with an and the were in a monocyte of Lp(a) isolated from a was higher than the Lp(a) from a low a to cells inhibited monocyte Lp(a) isolated from the MCP-1 to the low increased monocyte migration. The MCP-1 bound to the Lp(a) was to from the the of the and a necessary for monocyte migration. These results that MCP-1 carried by Lp(a) in human plasma is an LDL is a major risk factor in development of atherosclerosis C. Noels H. Atherosclerosis: current pathogenesis and therapeutic options.Nat. Med. 2011; 17: 1410-1422Crossref PubMed Scopus (1559) Google Scholar, C.K. Witztum J.L. Atherosclerosis. The road ahead.Cell. 2001; 104: 503-516Abstract Full Text Full Text PDF PubMed Scopus (2636) Google Scholar). that LDL oxidized in vivo and that oxidation the of LDL with vascular cells to studies of of OxLDL Y.I. P. L. R. K. A. A. C.J. X. et are associated by receptors of Res. 2011; PubMed Scopus Google Scholar, D. Witztum J.L. Oxidized lipoprotein and Biol. PubMed Scopus Google Scholar). These the of OxLDL in macrophages and vascular cells, and the activation of S. J.R. A.J. J.A. of oxidation in Res. PubMed Scopus Google Scholar, S. of macrophages in to 2011; PubMed Scopus Google Scholar). These of OxLDL are to the of oxidized and oxidized The that, in to oxidized OxLDL also a of such as the chemokine MCP-1. We demonstrate that in vitro OxLDL is of binding MCP-1 and a previously reported that in vitro OxLDL monocyte chemoattractant in to the of in the OxLDL S. D. modified low density a role in recruitment and of during Natl. Acad. Sci. USA. PubMed Scopus Google Scholar, S. D. a chemotactic factor for human and its role in Natl. Acad. Sci. USA. PubMed Scopus Google Scholar). MCP-1 binding to OxLDL results in a chemotactic on than that induced by OxLDL oxidized LDL is not to in in plasma but is in the wall D. Witztum J.L. Oxidized lipoprotein and Biol. PubMed Scopus Google Scholar). studies demonstrate that the major lipoprotein of OxPLs in human plasma, Lp(a), also binds MCP-1. with OxLDL, of MCP-1 to Lp(a) into a is the that endogenous MCP-1 is on Lp(a) in human plasma and that Lp(a) isolated from the plasma with a is a chemoattractant than the Lp(a) from low These results that Lp(a) in plasma may as a for MCP-1 and that the with its associated may the trafficking of to the vascular Lp(a) is in the human and a higher of Lp(a) with the of the clinical F. A. Leibundgut G. A. E. E. Curtiss L.K. Witztum J.L. et expression of and in and human coronary and atherosclerotic Lipid Res. Full Text Full Text PDF PubMed Scopus Google Scholar, G. R. P.C. Marcovina S.M. G. J.A. and in human coronary association with the of clinical Am. Coll. Cardiol. 1998; PubMed Scopus Google Scholar). into the vascular Lp(a) binds to of via its binding results in and of Lp(a) in the vascular wall S. J.L. as a risk factor of cardiovascular a for increased to its and Am. Coll. Cardiol. PubMed Scopus Google Scholar), where its may a of major and also may induce of macrophages M.J. X. J. M. R. Witztum J.L. Tsimikas S. et and lipoproteins in macrophages Full Text Full Text PDF PubMed Scopus Google Scholar). Lp(a)-associated MCP-1 may also to recruitment of to the lesion site and lesion progression. the Lp(a) of MCP-1 may added to the of of Lp(a) that a lipoprotein more than after 50 PubMed Scopus Google Scholar). the clinical that we was between levels and clinical We to studies to the role of in cardiovascular and the that clinical The of MCP-1 binding with OxLDL and Lp(a) is similar to that with GAGs and on GAGs and OxLDL are electronegative and the of basic Arg-18 and Lys-19 in MCP-1 with Ala of binding to GAGs or OxLDL. not all which to basic bind to GAGs (1Salanga C.L. Handel T.M. Chemokine oligomerization and interactions with receptors and glycosaminoglycans: the role of structural dynamics in function.Exp. Cell Res. 2011; 317: 590-601Crossref PubMed Scopus (132) Google Scholar). a to and chemokines on on the and to of and The interactions also chemokine from and and (1Salanga C.L. Handel T.M. Chemokine oligomerization and interactions with receptors and glycosaminoglycans: the role of structural dynamics in function.Exp. Cell Res. 2011; 317: 590-601Crossref PubMed Scopus (132) Google Scholar). in vitro the binding MCP-1 mutant which was also in in the in vivo is to recruit (7Proudfoot A.E. Handel T.M. Johnson Z. Lau E.K. LiWang P. Clark-Lewis I. Borlat F. Wells T.N. Kosco-Vilbois M.H. Glycosaminoglycan binding and oligomerization are essential for the in vivo activity of certain chemokines.Proc. Natl. Acad. Sci. USA. 2003; 100: 1885-1890Crossref PubMed Scopus (643) Google Scholar). These results the of binding for in vivo MCP-1 results that GAGs and Lp(a) for the MCP-1 binding to the of MCP-1 in vivo and studies demonstrate whether binding of MCP-1 to Lp(a) and OxLDL is important for the function of MCP-1 in the development of as in the chemokine (C-C motif) ligand 2 chemokine type 2 glycosaminoglycan lipoprotein(a) monoclonal antibody monocyte chemoattractant protein-1 myocardial infarction native LDL oxidized low density lipoprotein oxidized oxidized