Protein engineering and properties of human metalloproteinase and thrombospondin 1.
This work generated many truncated proteins and Glu(385) to Ala (E(385)/A) mutants of the human metalloproteinase and thrombospondin 1 (METH-1 or ADAMTS1) and specific antibodies. METH-1 was an active endopeptidase and both the metalloproteinase and the disintegrin/cysteine-rich domains were required for the proteinase activity. A point mutation at the zinc-binding site (E(385)/A) abolished the catalytic activity. METH-1 protein function may be modulated through proteolytic cleavage at multiple sites. One 135 kDa species had an NH(2)-terminal sequence of L(33)GRPSEEDEE. A species at 115 kDa and some other protein bands began with F(236)VSSHRYV(243), indicating that METH-1 proenzyme might be activated by a proprotein convertase such as furin by cleaving the R(235)-F(236) peptide bond. This cleavage was not an autocatalytic process since the E(385)/A mutants were also processed. Furthermore, a 52 kDa band with an NH(2)-terminal sequence of L(800)KEPLTIQV resulted from the digestion between the first and the second thrombospondin 1-like motifs in the spacer region of the extracellular matrix-binding domains.
Wei
Ping
P
Human Genome Sciences Inc., Rockville, MD 20850, USA.
Zhao
Yun-Ge
YG
Zhuang
Li
L
Hurst
Douglas R
DR
Ruben
Steve
S
Sang
Qing-Xiang Amy
QX
D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; D000409:Alanine; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D019483:Disintegrins; D018698:Glutamic Acid; D006801:Humans; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D015202:Protein Engineering; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000378:metabolism; D019700:Thrombospondin 1 / Q000737:chemistry / Q000235:genetics*
D019483:Disintegrins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D019700:Thrombospondin 1; D018698:Glutamic Acid; D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; C103859:ADAMTS1 protein, human; D008666:Metalloendopeptidases; D000409:Alanine