Purification and N-terminal sequence analysis of Streptomyces chromofuscus phospholipase D.
Partially purified commercial phospholipase D (PLD) was fractionated by dye-ligand affinity chromatography and nondenaturing polyacrylamide gel electrophoresis (PAGE). Active material migrated as three bands on SDS-PAGE. The two higher-abundance species were shown to have identical N-terminal sequences, while the third band was present in much smaller amounts and had a distinct sequence. Cloning Streptomyces chromofuscus PLD will allow the construction of stable transfectants of mast cell lines permitting regulated expression of PLD.
Dinh
T T
TT
Division of Allergy and Immunology, University of Texas Southwestern Medical Center, Dallas 75235-8859, USA.
McClure
G D
GD
Kennerly
D A
DA
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010739:Phospholipase D / Q000737:chemistry / Q000302:isolation & purification*; D013302:Streptomyces / Q000201:enzymology*
D001426:Bacterial Proteins; D010739:Phospholipase D