Isolation, characterization, and immunological detection of neutrophil-activating peptide 2: a proteolytic degradation product of platelet basic protein.
Neutrophil-activating peptide 2 (NAP-2), corresponding to platelet basic protein fragment 25-94, was prepared by chymotryptic digestion of its precursors, low affinity platelet factor 4 or beta-thromboglobulin, followed by purification by high performance liquid chromatography. NAP-2 (0.1-1.5 microns) caused the release of human granulocyte elastase from cytochalasin B-treated neutrophils in a dose-dependent manner. In the same system, beta-thromboglobulin, human platelet factor 4, S-pyridylethyl NAP-2, and platelet basic protein C-terminal fragment (77-94) were inactive, whereas platelet basic protein fragment 22-89 had low, but significant, activity. Sensitive immunological identification of NAP-2 based on nonequilibrium isoelectric focusing and immunoblotting is described.
Holt
J C
JC
Rhône-Poulenc Rorer Biotechnology, Inc., King of Prussia, Pennsylvania 19406.
Yan
Z Q
ZQ
Lu
W Q
WQ
Stewart
G J
GJ
Niewiarowski
S
S
D000595:Amino Acid Sequence; D001779:Blood Coagulation Factors / Q000378:metabolism; D018925:Chemokines; D002851:Chromatography, High Pressure Liquid; D003571:Cytochalasin B; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006494:Heparin Antagonists / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007525:Isoelectric Focusing; D019272:Leukocyte Elastase; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism; D010196:Pancreatic Elastase / Q000096:biosynthesis; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification; D010978:Platelet Factor 4; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D001620:beta-Thromboglobulin / Q000235:genetics / Q000378:metabolism
D001779:Blood Coagulation Factors; D018925:Chemokines; D006494:Heparin Antagonists; C061615:PPBP protein, human; D010455:Peptides; D011506:Proteins; D001620:beta-Thromboglobulin; D010978:Platelet Factor 4; D003571:Cytochalasin B; C009862:connective tissue-activating peptide; C018329:low affinity platelet factor 4; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase