Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Powerpoint - Blood Journal
Document related concepts
no text concepts found
Transcript
Human cathelicidin, hCAP-18, is processed to the antimicrobial peptide LL-37 by extracellular cleavage with proteinase 3 by Ole E. Sørensen, Per Follin, Anders H. Johnsen, Jero Calafat, G. Sandra Tjabringa, Pieter S. Hiemstra, and Niels Borregaard Blood Volume 97(12):3951-3959 June 15, 2001 ©2001 by American Society of Hematology Cleavage sites of cathelicidins.Cleavage sites between the cathelin part and the antimicrobial peptide of the bovine and porcine cathelicidins, cleaved by elastase, compared with the cleavage site of hCAP-18. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Electron microscopy of neutrophils from skin windows after phagocytosis of latex beads.(A) Cryosection incubated with anti–hCAP-18 and 10-nm protein A-gold. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Immunoblotting of cell lysates.Neutrophils were TCA-precipitated, and the precipitates were run on SDS-PAGE and analyzed by immunoblotting with anti–hCAP-18 antibodies. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Immunoblotting of exocytosed material.After stimulation, the neutrophils were pelleted and the supernatant containing the exocytosed material was analyzed by SDS-PAGE and immunoblotting with anti–hCAP-18 antibodies. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Immunoblotting of hCAP-18 after incubation with azurophil granule proteins.Purified hCAP-18 was incubated with azurophil granule proteins. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Immunoblotting with monoclonal anti–LL-37 antibody.All samples were run on SDS-PAGE followed by immunoblotting with monoclonal anti–LL-37 antibody. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Cleavage experiment with exocytosed material.Exocytosed material from ionomycin-stimulated neutrophils was depleted of endogenous hCAP-18 and subsequently incubated with purified hCAP-18 with or without protease inhibitors. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Differential inhibition of hCAP-18 cleavage by serine proteases from azurophil granules.Samples were run on SDS-PAGE followed by immunoblotting with anti–hCAP-18 antibodies. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Cleavage of hCAP-18 by exocytosed material after the immunoprecipitation of individual serine proteases.Endogenous hCAP-18 fragments were deleted from the exocytosed material. Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology Susceptibility of lipoprotein-bound hCAP-18 to cleavage by proteinase 3.Lipoprotein-bound hCAP-18 (lane a) was incubated with proteinase 3 (lane b). Ole E. Sørensen et al. Blood 2001;97:3951-3959 ©2001 by American Society of Hematology
