Activation of the protease from human adenovirus type 2 is accompanied by a conformational change that is dependent on cysteine-104.
Adenovirus codes for a protease the activity of which can be regulated in vitro by an 11 residue peptide (GVQSLKRRRCF) derived from another viral protein, pVI. Three cysteine residues, one in the activating peptide and two in the protease (C104 and C122), play a central role in both activation and catalysis. Expression of protease mutants in insect cells has shown that C104 is not essential for proteolytic activity. GVQSLKRRRCF also caused a concentration-dependent increase in tryptophan fluorescence of protease expressed in Escherichia coli that paralleled the increase in proteolytic activity, indicating that activation was accompanied by a conformational change. Tryptophan fluorescence of C104S was not increased by the addition of GVQSLKRRRCF, nor was the fluorescence of wild-type protease increased by the addition of the peptide analogues where cysteine is replaced by aspartic acid or serine, suggesting that C104 is involved in activation and C122 in catalysis.
Jones
S J
SJ
Division of Cell and Molecular Biology, School of Biological and Medical Sciences, University of St Andrews, UK.
Iqbal
M
M
Grierson
A W
AW
Kemp
G
G
D000260:Adenoviruses, Human / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D004279:DNA, Viral; D004789:Enzyme Activation; D006801:Humans; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology; D013329:Structure-Activity Relationship; D014764:Viral Proteins
D004279:DNA, Viral; D010750:Phosphoproteins; D011498:Protein Precursors; D014764:Viral Proteins; C085476:pTP protein, adenovirus; D003546:Cysteine Endopeptidases; C082284:L3 23K protein, adenovirus; D003545:Cysteine