Characterization, crystallization and preliminary X--ray diffraction analysis of acutohaemolysin, a haemolytic toxin from Agkistrodon acutus venom.
Acutohaemolysin, a phospholipase A(2) (PLA(2)) from the venom of the snake Agkistrodon acutus, has been isolated and purified to homogeneity by anion-exchange chromatography on a DEAE-Sepharose column followed by cation-exchange chromatography on a CM-Sepharose column. It is an alkaline protein with an isoelectric point of 10.5 and is comprised of a single polypeptide chain of 13 938 Da. Its N-terminal amino-acid sequence shows very high similarity to Lys49-type PLA(2) proteins from other snake venoms. Although its PLA(2) enzymatic activity is very low, acutohaemolysin has a strong indirect haemolytic activity and anticoagulant activity. Acutohaemolysin crystals with a diffraction limit of 1.60 A were obtained by the hanging-drop vapour-diffusion method. The crystals belong to the space group C2, with unit-cell parameters a = 45.30, b = 59.55, c = 46.13 A, beta = 117.69 degrees. The asymmetric unit contains one molecule.
Huang
Q Q
QQ
Laboratory of Structural Biology, University of Science and Technology of China, CAS, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China.
Zhu
X Y
XY
Li
N
N
Deng
W H
WH
Chen
T B
TB
Rao
P F
PF
Teng
M K
MK
Niu
L W
LW
D017836:Agkistrodon; D000595:Amino Acid Sequence; D000818:Animals; D002852:Chromatography, Ion Exchange; D003435:Crotalid Venoms / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D054501:Group II Phospholipases A2; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000737:chemistry* / Q000302:isolation & purification; D011487:Protein Conformation; D030162:Reptilian Proteins; D017386:Sequence Homology, Amino Acid
D003435:Crotalid Venoms; D030162:Reptilian Proteins; D010741:Phospholipases A; D054501:Group II Phospholipases A2; C414184:acutohemolysin, Agkistrodon acutus