Identification of an Epitope of α-Enolase (a Candidate Plasminogen Receptor) by Phage Display

Begoña Arza; Jordi Félez; Roser Lopez-Alemany; Lindsey A Miles; Pura Munñoz-Cánoves

doi:10.1055/s-0038-1657693

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1997; 78(03): 1097-1103
DOI: 10.1055/s-0038-1657693

Rapid Communication

Schattauer GmbH Stuttgart

Identification of an Epitope of α-Enolase (a Candidate Plasminogen Receptor) by Phage Display

Begoña Arza

¹The Institut de Recerca Oncòlogica (IRO), Hospital Duran i Reynals, Barcelona, Spain

,

Jordi Félez

¹The Institut de Recerca Oncòlogica (IRO), Hospital Duran i Reynals, Barcelona, Spain

,

Roser Lopez-Alemany

¹The Institut de Recerca Oncòlogica (IRO), Hospital Duran i Reynals, Barcelona, Spain

,

Lindsey A Miles

²The Scripps Research Institute, La Jolla, California, USA

,

Pura Munñoz-Cánoves

¹The Institut de Recerca Oncòlogica (IRO), Hospital Duran i Reynals, Barcelona, Spain

› Institutsangaben

Abstract

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Summary

α-enolase is an ubiquitous cytoplasmic glycolytic enzyme which also exhibits cell surface mediated functions and a structural role in the lens of some species. An α-enolase related molecule (a-ERM) is present on the surfaces of neutrophils, monocytes and monocytoid cells and has the capacity to specifically bind plasminogen, suggesting that a-ERM may function as a plasminogen receptor. We have generated a monoclonal antibody (mAB), 9C12, against a-ERM. This mAB reacted with both α-ERM and purified human α-enolase in Western blotting and in enzyme linked immunosorbent assays (ELISA). mAB 9C12 detected a cell surface associated molecule on human peripheral blood neutrophils and on U937 human monocytoid cells as assessed by fluorescence activated cell sorting (FACS) analyses. In addition, mAB 9C12 recognized an intracellular pool of a-enolase/a-ERM in permea bilized U937 cells. A phage display approach was employed to identify the a-enolase epitope recognized by mAB 9C12. Random fragments of 100-300 base pairs (bp), obtained from the full length human α-enolase cDNA, were cloned into the filamentous phage vector pComb3B, to generate a phage-displayed peptide library. Recombinant phages binding to mAB 9C12 were selected and their DNA inserts characterized by direct sequencing. All of the fragments which bound to mAB 9C12 en coded the common sequence DLDFKSPDDPSRYISP, spanning amino acids 257-272 of human α-enolase. This sequence is located within an external loop of the molecule. These data indicate that this sequence contains the epitope recognized by mAB 9C12 and is, therefore, exposed on the cell surface, further suggesting that a-enolase and a-ERM share common amino acid sequences.

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Referenzen

References
1 Wold F, Eno lase. in: The Enzymes. Boyer DP. (ed.). 03. ed. New York: Academic Press; 1971. pp 499-538
2 Bottalico LA, Kendrick NC, Keller A, Li Y, Tabas I. Cholesteryl ester loading of mouse peritoneal macrophages is associated with changes in the expression or modification of specific cellular proteins, including increase in an alpha-enolase isoform. Arteriosclerosis and Thrombosis 1993; 13: 264-275
3 Verma M, Kurl RN. Human lung enolase: Cloning and sequencing of cDNA and its inducibility with dexamethasone. Biochem Int 1993; 30: 293-303
4 Hawley SB, Miles LA. Characterization of a novel plasminogen binding protein associated with the U937 cell surface. Fibrinolysis 1996; 10: 32 (Abstract).
5 Piatigorski J. Lens crystallins. Innovation associated with changes in gene regulation. J Biol Chem 1992; 174: 5204-5210
6 Miles LA, Dahlberg CM, Plescia J, Felez J, Kato K, Plow EF. Role of Cell Surface Lysines in Plasminogen Binding to Cells: Identification of alfa Enolase as Candidate Plasminogen Receptor. Biochemistry 1991; 30: 1682-1691
7 Fukao H, Hagiya Y, Nonaka T, Okada K, Matsuo O. Analysis of binding protein for tissue-type plasminogen activator in human endothelial cells. Biochem Biophys Res Commun 1992; 187: 956-962
8 Nakajima K, Hamanoue M, Takemoto N, Hattori T, Kato K, Kohsaka S. Plasminogen binds specifically to alpha-enolase on rat neuronal plasma membrane. Journal of Neurochemistry 1994; 63 (06) 2048-2056
9 Redlitz A, Fowler BJ, Plow EF, Miles LA. The role of an enolase-related molecule in plasminogen binding to cells. Eur J Biochem 1995; 227: 407-415
10 Andronicos NM, Ranson M, Bognacki J, Baker MS. The human ENOl gene product (recombinant human alpha-enolase) displays characteristics required for a plasminogen binding protein. Biochim Biophys Acta 1997; 1337: 27-39
11 Felez J, Miles LA, Fabregas P, Jardi M, Plow EF, Lijnen HR. Characteriza tion of cellular binding sites and interactive regions within reactants re quired for enhancement of plasminogen activation by tPA on the surface of leukocytic cells. Thromb Haemost 1996; 76: 577-585
12 Giallongo A, Feo S, Moore R, Croce CM, Showe LC. Molecular cloning and nucleotide sequence of a full-length cDNA for human alpha-enolase. Proc Natl Acad Sci USA 1986; 83: 6741-6745
13 Hoess RH. Phage display of peptides and protein domains. Curr Opin Struct Biol 1993; 03: 572-579
14 Barbas CF. Recent advances in phage display. Curr Opin Biotechnol 1993; 04: 526-530
15 Barbas CF, Barbas SM. Filamentous phage display. Fibrinolysis 1994; 08 (01) 245-252
16 Todd III FR, Bamathan ES, Bohuslav J, Chapman HA, Cohen RL, Felez J, Howell A, Johnson JG, Knapp W, Kramer M, Miles LA, Nykjaer A, Ralfkiaer E, Shuren E. M15 Cluster Report: CD87. in: Leucocyte Typing V: White Cell Differentiation Antigens. Schloosman SF, Boumsell L, Gilks W, Harlan JM, Kishimoto T, Morimoto C, Ritz J, Shaw S, Silverstein RL, Springer TA, Tedder TF, Todd III FR. (eds.). Oxford: Oxford University Press; 1995. pp 932-939
17 Plow EF, Freaney DE, Plescia J, Miles LA. The plasminogen system and cell surfaces: evidence for plasminogen and urokinase receptors on the same cell type. J Cell Biol 1986; 103: 2411-2420
18 Felez J, Miles LA, Plescia J, Plow EF. Regulation of plasminogen recep tors on human monocytes and monocytoid cells. J Cell Biol 1990; 111: 1673-1683
19 Felez J, Chanquia CJ, Fabregas P, Plow EF, Miles LA. Competition between plasminogen and t-PA for cellular binding sites. Blood 1993; 82: 2433-2441
20 Laemmli UK. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 1970; 227: 280-282
21 van ZonneveldA, van denBerg, van MeijerM, Pannekoek H. Identification of functional interaction sites on proteins using bacteriphage displayed random epitope libraries. Gene 1995; 167: 49-52
22 Barbas CF, Kang AS, Lemer RA, Benkovic SJ. Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc Natl Acad Sci USA 1991; 88: 7978-7982
23 Belin D, Vassalli J, Combepine C, Godeau F, Nagamine Y, Reich E, Kocher HP, Duvoisin RM. Cloning, nucleotide sequencing and expression of cDNAs encoding mouse urokinase-type plasminogen activator. Euro pean Journal Biochemistry 1985; 148: 225-232
24 Barbas III CF, Lemer RA. Combinatorial immunoglobulin libraries on the surface of phage (Phabs); rapid selection of antigen-specific Fabs. Methods Enzymol 1991; 02: 119-124
25 Miles LA, Plow EF. Receptor mediated binding of the fibrinolytic components, plasminogen and urokinase, to peripheral blood cells. Thromb Haemost 1987; 58: 936-942
26 Jardi M, Cuervo JH, Fabregas P, Rodriguez-Lambies N, Felez J. Monoclo nal antibodies to different fibrinolytic components. Biol Clin Hematol 1995; 17: 18-27
27 Smith GP, Scott JK. Libraries of peptides and proteins displayed on fila mentous phage. Methods Enzymol 1993; 217: 228-232
28 Hajjar KA, Nachman RL. Endothelial cell-mediated conversion of glu-plas minogeri to lys-plasminogen. J Clin Invest 1988; 82: 1769-1778
29 Plow EF, Felez J, Miles LA. Cellular regulation of fibrinolysis. Thromb Haemost 1991; 66: 32-36
30 Ellis V. Functional analysis of the cellular receptor for urokinase in plasminogen activation. J Biol Chem 1996; 271: 14779-14784
31 Plow EF, Miles LA. Plasminogen receptors in the mediation of pericellular proteolysis. Cell Differ Dev 1990; 32: 293-8
32 Parkkinen J, Rauvala H. Interactions of plasminogen and tissue plas minogen activator (t-PA) with amphoterin. Enhancement of t-PA-cata lyzed plasminogen activation by amphoterin. J Biol Chem 1991; 266: 16730-16735
33 Miles LA. Regulation of fibrinolysis and proteolysis by plasminogen receptors. Biol Clin Hematol 1993; 15: 107-115
34 Dudani AK, Cummings C, Hashemi S, Ganz PR. Isolation of a novel 45 kDa plasminogen receptor from human endothelial cells. Thromb Res 1993; 69: 185-196
35 Hajjar KA, Jacovina AT. Molecular cloning of a endothelial cell tissue plasminogen activator/plasminogen receptor: annexin II-related protein. Thromb Haemost 1993; 69: 989-A-1601
36 Lopez-Alemany R, Correc P, Camoin L, Burtin P. Purification of the plas min receptor from human carcinoma cells and comparison to alpha-enolase. Thromb Res 1994; 75 (04) 371-381
37 Hajjar KA, Jacovina AT, Chacko J. An endothelial cell receptor for plasmi nogen/tissue plasminogen activator. I. Identity with annexin II. J Biol Chem 1994; 269: 21191-21197
38 Cesarman GM, Guevara CA, Hajjar KA. An endothelial cell receptor for plasminogen/tissue plasminogen activator (t-PA). II. Annexin mediated en hancement of t-PA dependent plasminogen activation. J Biol Chem 1994; 269: 21198-21203
39 Dudani AK, Pluskota A, Ganz PR. Interaction of tissue plasminogen activa tor with a human endothelial cell 45-kilodalton plasminogen receptor. Biochem Cell Biol 1994; 72: 126-131
40 Hembrough TA, Vasudevan J, Allietta MM, Glass II FW, Gonias SL. A cytokeratin 8-like protein with plasminogen-binding activity is present on the external surfaces of hepatocytes, HepG2 cells and breast carcinoma cell lines. J Cell Scien 1995; 108: 1071-1082
41 Winram SB, Lottenberg R. The plasmin-binding protein Plr of group A streptococci is identified as glyceraldehyde-3-phosphate dehydrogenase. Microbiology 1996; 142: 2311-2320
42 Hajjar KA, Guevara CA, Lev E, Dowling K, Chacko J. Interaction of the fibrinolytic receptor, annexin II, with the endothelial cell surface – Essential role of endonexin repeat. J Biol Chem 1996; 271: 21652-21659
43 Lopez-Alemany R, Longstaff C, Fabregas P, Jardi M, Merton E, Félez J. Inhibition of plasmin generation on cell surfaces by monoclonal antibodies against a 55-60 kDa plasminogen receptor. Fibrinolysis 1996; 10: 16 (Abstract).
44 Miles LA, Dahlberg CM, Levin EG, Plow EF. Gangliosides interact directly with plasminogen and urokinase and may mediate binding of these fibrinolytic components to cells. Biochemistry 1989; 28: 9337-9343
45 von HeijneG, Liljestrom P, Mikus P, Anderson H, Ny T. The efficiency of the uncleaved secretion signal in the plasminogen activator inhibitor type 2 protein can be enhanced by point mutations that increase its hydrophobic ity. J Biol Chem 1991; 266: 15240-15243
46 Lebioda L, Stec B, Brewer JM. The structure of yeast enolase at 2.25-A resolution. An 8-fold beta + alpha-barrel with a novel beta beta alpha alpha (beta alpha) 6 topology. J Biol Chem 1989; 264: 3685-3693
47 Peitsch MC. ProMod and Swiss-Model: Internet-based tools for auto mated comparative protein modelling. Biochem Soc Trans 1996; 24: 274-279
48 Cali L, Feo S, Oliva D, Giallongo A. Nucleotide sequence of a cDNA encoding the human muscle-specific enolase (MSE). Nucleic Acids Res 1990; 18: 1893