title: A novel CD44-binding peptide from the pro-matrix metalloproteinase-9 hemopexin domain impairs adhesion and migration of chronic lymphocytic leukemia (CLL) cells.
abstract: (pro)MMP-9 binds to CLL cells through the PEX9 domain and contributes to CLL progression. To biochemically characterize this interaction and identify potential therapeutic targets, we prepared GST-PEX9 forms containing structural blades B1B2 or B3B4. We recently described a sequence in blade B4 (P3 sequence) that bound α4β1 integrin and partially impaired cell adhesion and migration. We have now studied the possible contribution of the B1B2 region to cell interaction with PEX9. CLL cells bound to GST-B1B2 and CD44 was the primary receptor. GST-B1B2 inhibited CLL cell migration as effectively as GST-B3B4. Overlapping synthetic peptides spanning the B1B2 region identified the sequence FDAIAEIGNQLYLFKDGKYW, present in B1 and contained in peptide P6, as the most effective site. P6 inhibited cell adhesion to PEX9 in a dose-dependent manner and with an IC50 value of 90 μM. P6 also inhibited cell adhesion to hyaluronan but had no effect on adhesion to VCAM-1 (α4β1 integrin ligand), confirming its specific interaction with CD44. Spatial localization analyses mapped P6 to the central cavity of PEX9, in close proximity to the previously identified P3 sequence. Both P6 and P3 equally impaired cell adhesion to (pro)MMP-9. Moreover, P6 synergistically cooperated with P3, resulting in complete inhibition of CLL cell binding to PEX9, chemotaxis, and transendothelial migration. Thus, P6 is a novel sequence in PEX9 involved in cell-PEX9/(pro)MMP-9 binding by interacting with CD44. Targeting both sites, P6 and P3, should efficiently prevent (pro)MMP-9 binding to CLL cells and its pathological consequences.
authors.affiliation: From the Department of Cellular and Molecular Medicine, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, Spain.
authors.forename: Estefanía
authors.initials: E
authors.lastname: Ugarte-Berzal
authors.affiliation: From the Department of Cellular and Molecular Medicine, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, Spain.
authors.forename: Elvira
authors.initials: E
authors.lastname: Bailón
authors.affiliation: From the Department of Cellular and Molecular Medicine, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Amigo-Jiménez
authors.affiliation: the Proteomics Facility, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, 28049 Madrid, Spain, and.
authors.forename: Juan Pablo
authors.initials: JP
authors.lastname: Albar
authors.affiliation: Servicio de Hematología, Hospital Universitario Puerta de Hierro, 28222 Madrid, Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: García-Marco
authors.affiliation: From the Department of Cellular and Molecular Medicine, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, Spain, agarciapardo@cib.csic.es.
authors.forename: Angeles
authors.initials: A
authors.lastname: García-Pardo
mesh_terms: D000368:Aged; D000595:Amino Acid Sequence; D002448:Cell Adhesion / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D018450:Disease Progression; D015195:Drug Design; D004792:Enzyme Precursors / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006466:Hemopexin / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry / Q000378:metabolism*; D008875:Middle Aged; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary
chemical_list: C497483:CD44 protein, human; D004792:Enzyme Precursors; D018960:Hyaluronan Receptors; D010455:Peptides; D006466:Hemopexin; C120487:pro-matrix metalloproteinase 9; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9
keywords: CD44; Cell Adhesion; Cell Migration; Leukemia; Metalloprotease
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