Purification and N-terminal amino acid sequence of sheep neutrophil cathepsin G and elastase.
Sheep cathepsin G (CG) and neutrophil elastase (NE) were isolated from a crude leukocyte membrane preparation by elastin-Sepharose 4B and CM-Sepharose 4B chromatography, followed by native preparative PAGE. The N-termini of CG and NE were sequenced to 24 and 20 residues, showing 96 and 85% identity with human CG and NE, respectively. During SDS-PAGE, sheep CG and NE migrated parallel to human CG and NE and have apparent molecular masses of 28 and 26 kDa, respectively. Following incubation of sheep CG and NE with human alpha(1)-antichymotrypsin and alpha(1)-proteinase inhibitor, complexes with apparent molecular masses of 89 and 81 kDa respectively were observed by SDS-PAGE. Polyclonal antibodies to human CG and NE cross-reacted with purified sheep CG and NE, respectively. These results indicate that sheep neutrophils contain CG and elastase that are analogous to human CG and NE in terms of molecular mass, reactivity with endogenous inhibitors, immunocross-reactivity, and N-terminal sequence.
Department of Respiratory Medicine, Charing Cross Hospital, National Heart and Lung Institute, Imperial College School of Medicine, London, W6 8RF, United Kingdom.
R
R
Mistry
P D
PD
Snashall
N
N
Totty
A
A
Guz
T D
TD
Tetley
D000595:Amino Acid Sequence; D000818:Animals; D056649:Cathepsin G; D002403:Cathepsins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D003429:Cross Reactions; D006801:Humans; D007120:Immunochemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009504:Neutrophils / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases; D012756:Sheep; D013045:Species Specificity
D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D010196:Pancreatic Elastase