title: Functional region of mouse heat shock protein 72 for its binding to lymphoid neoplastic P388D1 cells.
abstract: Extracellular heat shock proteins have been reported to participate in both innate and adaptive immune responses. We have found that recombinant mouse inducible heat shock protein 72 (Hsp72) bound to lymphoid neoplastic P388D1 cells. In the present study, we examined which region of mouse Hsp72 interacted with this cell line by using truncated variants that are sequentially lacking sections of the C-terminal region. The full-length mouse Hsp72 specifically bound to P388D1 cells, but not to mastocytoma P815 cells. Deletion of the C-terminal tail portion of mouse Hsp72 markedly decreased the binding to P388D1 cells and the sequential truncation of the C-terminal helical region led to a loss of binding activity. Specific binding was not observed for either the variant with a minimal substrate-binding structure or the ATP-binding domain alone. On the other hand, two truncated variants lacking the ATP-binding domain significantly bound to P388D1 cells. However, the variant lacking the substrate-binding domain did not show any binding to this cell line. These results suggest that the activity to bind P388D1 cells is attributable to the C-terminal region of mouse Hsp72 in combination with the substrate-binding domain. Interestingly, the binding of mouse Hsp72 to P388D1 cells was competed by the variant with the C-terminal flexible tail sequence, but not by the variant without that sequence. These competitive experiments imply that there may be at least two membrane receptors on P388D1 cells and also that both receptors may recognize the various structures in the C-terminal region of the Hsp70 family for regulation of innate immunity.
authors.affiliation: Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Goka-sho, Uji, Kyoto 611-011, Japan.
authors.forename: Michiko
authors.initials: M
authors.lastname: Ohno
authors.forename: Naofumi
authors.initials: N
authors.lastname: Kitabatake
authors.forename: Fumito
authors.initials: F
authors.lastname: Tani
mesh_terms: D000818:Animals; D019921:Biotinylation; D050884:HSP72 Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D008223:Lymphoma / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D050884:HSP72 Heat-Shock Proteins; D050505:Mutant Proteins; D011994:Recombinant Proteins