Purification and amino acid sequence of basic protein I, a lysine-49-phospholipase A2 with low activity, from the venom of Trimeresurus flavoviridis (Habu snake).
A basic protein (pI 10.2), named basic protein I, was purified to homogeneity from the venom of Trimeresurus flavoviridis (Habu snake) after four chromatographic steps. The amino acid sequence of this protein was determined by sequencing the S-pyridylethylated derivative of the protein and its peptides produced by chemical (cyanogen bromide and formic acid) and enzymatic (chymotrypsin, Achromobacter protease I, and Staphylococcus aureus V8 protease) cleavages. The protein consisted of 122 amino acid residues and was similar in sequence to phospholipases A2 from the venoms of crotalid and viperid snakes. A most striking feature of this protein is that aspartic acid at the 49th position common in phospholipases A2 is replaced by lysine. When the protein acted on 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphorylcholine, oleic acid was preferentially released, indicating that the protein has phospholipase A2 activity. Its molar activity toward 1,2-dilauroyl-sn-glycero-3-phosphorylcholine, however, was 1.5% that of T. flavoviridis phospholipase A2 isolated previously. The fact that both affinity to Ca2+ and reactivity to p-bromophenacyl bromide of basic protein I are approximately one order of magnitude lower than those of T. flavoviridis phospholipase A2 might explain the low activity of basic protein I.
Yoshizumi
K
K
Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka, Japan.
Liu
S Y
SY
Miyata
T
T
Saita
S
S
Ohno
M
M
Iwanaga
S
S
Kihara
H
H
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms / Q000032:analysis* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006639:Histidine / Q000032:analysis; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis; D010713:Phosphatidylcholines / Q000032:analysis; D010740:Phospholipases / Q000032:analysis*; D010741:Phospholipases A / Q000032:analysis* / Q000302:isolation & purification; D054467:Phospholipases A2; D011506:Proteins; D013386:Succinates / Q000032:analysis; D013438:Sulfhydryl Compounds / Q000032:analysis
D000596:Amino Acids; D002241:Carbohydrates; D003435:Crotalid Venoms; D010455:Peptides; D010713:Phosphatidylcholines; D011506:Proteins; D013386:Succinates; D013438:Sulfhydryl Compounds; C063854:basic protein I; D006639:Histidine; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2