曼陀罗
生物
生物化学
半胱氨酸蛋白酶
分子生物学
蛋白酵素
丝氨酸蛋白酶
半胱氨酸
蛋白酶
互补DNA
肽序列
胰蛋白酶
酶
基因
植物
幼虫
作者
Takayuki Miyaji,Yoshiaki Kouzuma,Jun Yaguchi,Rika Matsumoto,Michael R. Kanost,Karl J. Kramer,Masami Yonekura
标识
DOI:10.1016/j.ibmb.2007.05.003
摘要
A cysteine protease inhibitor (CPI) with an apparent molecular mass of 11.5 kDa was purified from larval hemolymph of the tobacco hornworm (Manduca sexta) by gel filtration on Sephadex G-50 followed by hydrophobic and ion-exchange column chromatographies. The purified cysteine proteinase inhibitor, denoted as MsCPI, strongly inhibited the plant cysteine protease, papain, with a Ki value of 5.5×10−9 M. Nucleotide sequence analysis of a partial cDNA encoding MsCPI indicated that MsCPI consists of 105 amino acid residues in a sequence that is similar to sarcocystatin A from Sarcophaga peregrina. However, northern blotting and PCR analyses using the specific primers of MsCPI suggested that the mRNA encoding MsCPI had a size of more than 12 kilobases, which included at least six tandemly repeated MsCPI segments. MsCPI was expressed in Escherichia coli and the recombinant protein effectively inhibited cysteine proteases from plants as well as from animals such as cathepsins B (Ki, 6.8 nM), H (3.0 nM), and L (0.87 nM). There was no inhibition exhibited toward trypsin, chymotrypsin, subtilisin, pepsin or themolysin.
科研通智能强力驱动
Strongly Powered by AbleSci AI