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Acidic pH binding sites for 2M * are also responsible for binding transferrin, lactoferrin and apoE at acidic pH in cells, we first examined the effects of these proteins on 125I- 2M * binding to Mv1Lu, MEF, and PEA-13 cells. Cells were incubated with 2 nM 125I- 2M * in the presence and absence of 10 M transferrin, lactoferrin, or apoE at pH 5.5 or pH 7.4 for comparison ; . After 2.5 h at 0 C, the specific binding of 125I- 2M * to cells was determined. At 10 M, all of these proteins completely blocked the specific binding at pH 5.5 ; of 125I- 2M * in Mv1Lu, MEF, and PEA-13 cells data not shown ; . In contrast, none of these proteins had a significant effect on 125I- 2M * binding at pH 7.4 ; to Mv1Lu and MEF cells, which is mediated by LRP-1 data not shown ; . Lactoferrin and 2M * bind to distinct sites of LRP-1 and do not compete with each other for binding to LRP-1 10, 12 ; . These results suggest that transferrin, lactoferrin and apoE may bind to the same acidic pH binding sites as 2M * does. Alternatively, the acidic pH binding sites for these molecules may be different but overlapping. To further define the ligand specificity of the acidic pH binding sites, the effects of various concentrations of transferrin, lactoferrin, -globulin, Pseudomonas exotoxin A also a ligand of LRP-1 ; 36 ; , or LDL on 125I- 2M * binding at pH 5 Mv1Lu cells were determined. As shown in Fig. 4, A and B, increasing concentrations.
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Devost NC, Choy FYM. 2000 ; Mutation analysis of Gaucher disease using dot-blood samples on FTA R filter paper. J Med Genet, 94: 417-420. Di Natale P, Balzano N, Esposito S, Villani G 1998 ; Identification of molecular defects in Italian Sanfilippo A patients including 13 novel mutations. Hum Mut, 11: 313320. Di Natale P, Vanacore B, Daniele A, Esposito S 2001 ; Heparan N-sulfatase: In vitro mutagenesis of potential Nglycosylation sites. Biochem Biophys Res Commun, 280: 1251-1257. Di Natale P, Villani GRD, Di Domenico C, Daniele A, Vici CD, Bartuli A. 2003 ; Analysis of Sanfilippo A gene mutations in a large pedigree. Clin Genet, 63: 314-318. Daz A, Montfort M, Cormand B, Zeng BJ, Pastores GM, Chabs A, Vilageliu L, i col. 1999 ; Gaucher disease: The N370S mutation in Ashkenazi Jewish and Spanish patients has a common origin and arose several thousand years ago. J Hum Genet, 64: 1233-1238. Daz A, Montfort M, Cormand B, Zeng BJ, Pastores GM, Chabs A, Vilageliu L, i col. 2000 ; On the age of the most prevalent Gaucher disease-causing mutation, N370S. J Hum Genet, 66: 2014-2015. Daz-Font A, Cormand B, Blanco M, Chamoles N, Chabs A, Grinberg D, Vilageliu L. 2003a ; Gene rearrangements in the glucocerebrosidase-metaxin region giving rise to disease-causing mutations and polymorphisms. Analysis of 25 RecNciI alleles in Gaucher disease patients. Hum Genet, 112: 426-429. Daz-Font A, Cormand B, Chabs A, Vilageliu L, Grinberg D. 2003b ; Unsuccessful chimeraplast strategy for the correction of a mutation causing Gaucher disease. Blood Cell Mol Dis, 31: 183-186. Dierks T, Schmidt B, von Figura K. 1997 ; Conversion of cysteine to formylglycine: a protein modification in the endoplasmatic reticulum. Proc Natl Acad Sci USA ; , 94: 11963-11968. Dierks T, Schmidt B, Borissenko LV, Peng J, Preusser A, Mariappan M, von Figura K. 2003 ; Multiple sulfatase deficiency is caused by mutations in de the gene encoding the human Ca-formylglycine generating enzyme. Cell, 113: 435-444. Doll RF, Bruce A, Smith FI. 1995 ; Regulation of the human acid beta-glucosidase promoter in multiple cell types. Bba-Gene Struct Express, 1261: 57-67.
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Activase TNKase Sales of our two cardiovascular products, Activase and TNKase, were 6.2 million in 2000, a decrease of 13% from 1999. TNKase received FDA approval in early June 2000 and was launched in late June 2000. The decrease from the prior year was due primarily to increased competition from Centocor, Inc.'s Retavase reteplase ; , and a decline in the overall size of the thrombolytic market as a result of increasing use of mechanical reperfusion as well as early intervention with other therapies in the treatment of acute myocardial infarction. In 1999, sales of Activase were 6.0 million, an increase of 11% from 1998. This increase was largely due to the usage of Activase in peripheral vascular occlusive disease in lieu of another company's thrombolytic that was unavailable. This increase was offset in part by a continued decline in the overall size of the thrombolytic therapy market due to increasing use of mechanical reperfusion and competition from Centocor's Retavase. Growth Hormone and metolazone.
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