Peptide substrate cleavage specificity of the human cytomegalovirus protease.
The human cytomegalovirus UL80 gene encodes an 80-kDa precursor polyprotein whose N-terminal 256-amino acid domain is a protease. This enzyme cleaves a specific peptide bond that results in its own release from the precursor, as well as a peptide bond near the C terminus of the viral assembly protein. The latter cleavage is apparently required for encapsidation of the viral genomic DNA and maturation of the viral capsid. A series of peptide substrates, representing the assembly protein cleavage site, was used to study the enzyme's substrate requirements and specificity. It was found that efficient cleavage minimally required the amino acid residues spanning the P4 to P4' positions. Substitution at any of these residues adversely affected the reaction. Conservation of the hydrophobic residues at P3 and P4 was essential. In addition, cleavage of a peptide representing the protease domain release site was reduced almost 100-fold relative to cleavage of the assembly protein maturation site peptide substrate.
Sardana
V V
VV
Department of Antiviral Research, Merck Research Laboratories, West Point, Pennsylvania 19486-0004.
Wolfgang
J A
JA
Veloski
C A
CA
Long
W J
WJ
LeGrow
K
K
Wolanski
B
B
Emini
E A
EA
LaFemina
R L
RL
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000201:enzymology* / Q000235:genetics; D017931:DNA Primers; D004279:DNA, Viral / Q000378:metabolism*; D010450:Endopeptidases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D005814:Genes, Viral; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D014764:Viral Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*
D017931:DNA Primers; D004279:DNA, Viral; D009842:Oligopeptides; D011994:Recombinant Proteins; D014764:Viral Proteins; D010450:Endopeptidases; C087225:human cytomegalovirus protease UL80