Purification and characterization of hementin, a fibrinogenolytic protease from the leech Haementeria ghilianii.
The fibrinogenolytic enzyme hementin, present in extracts of the posterior salivary glands of the giant leech Haementeria ghilianii, was isolated by ultrafiltration, high-performance ion-exchange chromatography and subsequent reversed-phase liquid chromatography. Approximately 100 micrograms (1 nmol) of hementin, present at less than 0.5% in the crude leech salivary extract, was brought to about 90% purity in three steps. Hementin migrated at an Mr of about 73,000 on non-reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and at 82,000 on reducing SDS-PAGE. The amino terminal sequence was determined to be TTLTE-PEPDL. The amino terminal sequences of two inactive proteins that partially coeluted with hementin in the first chromatographic step were also determined.
Smith Kline & French Laboratories, King of Prussia, PA 19406-0939.
J K
JK
Swadesh
I Y
IY
Huang
A Z
AZ
Budzynski
D000595:Amino Acid Sequence; D000818:Animals; D000925:Anticoagulants / Q000032:analysis / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D007865:Leeches / Q000032:analysis*; D008666:Metalloendopeptidases / Q000032:analysis / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000302:isolation & purification; D012469:Salivary Glands / Q000032:analysis
D000925:Anticoagulants; D010447:Peptide Hydrolases; D008666:Metalloendopeptidases; C040192:hementin