Cloning and expression in Pichia pastoris of metalloprotease domain of ADAM 9 catalytically active against fibronectin.
ADAM 9 is a member of the cellular metalloprotease/disintegrin/cysteine-rich (MDC) gene family, related to soluble snake venom metalloproteases (SVMP). ADAMs may play important roles in cell-cell fusion, cell-matrix interaction, and other cellular functions. To investigate catalytic activity of human ADAM 9 we have cloned and expressed the metalloprotease domain of human ADAM 9 in Pichia pastoris. The recombinant protein was purified in a three-step purification procedure and activity was detected against gelatin, beta-casein, and fibronectin. In addition we identified five normal and cancer cell lines expressing mRNA of human ADAM 9.
Schwettmann
L
L
Department of Biochemistry I, Faculty of Chemistry, University of Bielefeld, 33615 Bielefeld, Germany.
Tschesche
H
H
D051722:ADAM Proteins; D001943:Breast Neoplasms; D002384:Catalysis; D003001:Cloning, Molecular / Q000379:methods; D019483:Disintegrins; D004926:Escherichia coli; D005260:Female; D005353:Fibronectins / Q000378:metabolism*; D015723:Gene Library; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D009124:Muscle Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D019483:Disintegrins; D005353:Fibronectins; D008565:Membrane Proteins; D009124:Muscle Proteins; D011994:Recombinant Proteins; D051722:ADAM Proteins; C098339:ADAM9 protein, human; D008666:Metalloendopeptidases