Isolation and characterization of a hyaluronidase from the venom of Chinese red scorpion Buthus martensi.
A hyaluronidase, named BmHYA1, was purified from the venom of Chinese red scorpion (Buthus martensi), using successive chromatography. The homogeneity of BmHYA1 was confirmed by SDS-PAGE and MALDI-TOF mass spectrometry. The molecular mass of BmHYA1 was 48,696 Da determined by MALDI-TOF MS. The optimal temperature and pH of BmHYA1 were 50 degrees C and pH 4.5, respectively. It could be inhibited by DTT, Cu(2+), Fe(3+) or heparin, but not Mg(2+), Ca(2+), reduced glutathione, l-cysteine or EDTA. The sequence of thirty N-terminal amino acids of BmHYA1 was obtained by Edman degradation, as TSADF KVVWE VPSIM CSKKF KICVT DLLTS; but no similarity was found to other venom hyaluronidases. Further, BmHYA1 can hydrolyze hyaluronan into relatively smaller oligosaccharides and modulate the expression of CD44 variant in the breast cancer cell line MDA-MB-231.
Feng
Luo
L
Venom and Toxin Research Programme, Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, 117597 Singapore.
Gao
Rong
R
Gopalakrishnakone
Ponnampalam
P
D000818:Animals; D001943:Breast Neoplasms; D045744:Cell Line, Tumor; D004795:Enzyme Stability; D006801:Humans; D018960:Hyaluronan Receptors / Q000096:biosynthesis; D006820:Hyaluronic Acid / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D012604:Scorpion Venoms / Q000037:antagonists & inhibitors / Q000201:enzymology* / Q000302:isolation & purification / Q000378:metabolism; D012605:Scorpions; D013696:Temperature
C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D012604:Scorpion Venoms; D006820:Hyaluronic Acid; C532432:BmHYA1 protein, Buthus martensi; D006821:Hyaluronoglucosaminidase