[Expression, purification and identification of human matrix metalloproteinase-2].
The expression sequence of matrix metalloproteinase-2 (MMP-2) has been obtained by PCR amplifying, restriction enzyme cut and sequencing analysis demonstrate that the sequence is correct. The recombinant expression plasmid pPIC9/MMP-2 containing MMP-2 is constructed and transformed the yeast Pichia pastoris. Recombinant matrix metalloproteinase-2 protein was expressed in Pichia pastoris in great deal after induction by methanol. The purity of the recombinant MMP-2 filtrated through Sephacryl S-200 reached to electrophoresis purity. With the ability to degrade gelatin and IV type collagen, recombinant MMP-2 has the similar substrate specificity with natural MMP-2. The recombinant MMP-2 with 50 kD molecular weight is smaller than natural MMP-2, which suggested degradation occurred to it.
Ding
X Y
XY
dingxiuyun@mailexcite.com
Li
C H
CH
Sun
L Y
LY
Wang
H L
HL
D002847:Chromatography, Agarose; D024062:Collagen Type V / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000432:Methanol / Q000494:pharmacology; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D013379:Substrate Specificity
D024062:Collagen Type V; D011994:Recombinant Proteins; D005780:Gelatin; D020778:Matrix Metalloproteinase 2; D000432:Methanol