3.4.21.97 A133V I-site mutant 95603 3.4.21.97 A134Q oligomerization at high protein concentration 95600 3.4.21.97 A143Q all enzyme samples used in this experiments contain an additional mutation, A143Q. The mutation disables one of the internal cleavage sites but has little effects on the kinetic properties of the enzyme 653320 3.4.21.97 A143Q/D217N the ratio of turnover number to Km-value is 11% of that for mutant enzyme A143Q 653320 3.4.21.97 A143Q/D227N the ratio of turnover number to Km-value is 1273fold lower than that for mutant A143Q 653320 3.4.21.97 A143Q/E31R the ratio of turnover number to Km-value is 44.6fold lower than that for the wild-typelike mutant A143Q 650227 3.4.21.97 A143Q/E31S the ratio of turnover number to Km-value is 4.8fold lower than that for the wild-typelike mutant A143Q 650227 3.4.21.97 A143Q/H157A the ratio of turnover number to Km-value is 22fold lower than that of the wild-typelike mutant enzyme A143Q 650071 3.4.21.97 A143Q/H157E the ratio of turnover number to Km-value is 11.7fold lower than that of the wild-typelike mutant enzyme A143Q 650071 3.4.21.97 A143Q/H157Q the ratio of turnover number to Km-value is 19fold lower than that of the wild-typelike mutant enzyme A143Q 650071 3.4.21.97 A143Q/L229R mutant enzyme A143Q/L229R shows approximately a 20% decrease in helical content relative to the mutant enzyme A143Q, mutant enzyme A143Q/L229R has significant lower thermal stability than the mutant enzyme A143Q 653320 3.4.21.97 A143Q/L229R mutation has little effect on the stability of the dimer, significant lower thermal stability than the mutant enzyme A143Q. The ratio of turnover number to Km-value is 1826fold lower than that for mutant enzyme A143Q 653320 3.4.21.97 A143Q/R137E the ratio of turnover number to Km-value is 6.3fold lower than that for the wild-typelike mutant A143Q 650227 3.4.21.97 A143Q/R232H mutant enzyme A143Q/R232H essentially maintains wild-type catalytic activity, shows little change in the CD spectra compared to mutant A143Q enzyme, thermal stability is similar to that of mutant A143Q enzyme. The ratio of turnover number to Km-value is 5.1fold lower than that for mutant enzyme A143Q 653320 3.4.21.97 A143Q/S225Y mutant enzyme with large conformational changes with 40% lower helical content relative to the mutant enzyme A143Q, mutant dimer has similar stability to the mutant A143Q dimer. Many of the conformational differences between the mutant A143Q and the mutant enzyme A143Q/S225Y are likely the direct result of the mutation itself. alphaF helix tilts away from the wild type dimer two-fold axis in the S225Y mutant, creating a space near the two-fold axis to accomodate the new Tyr side chain. The S225Y mutation is the likely trigger for the change in the monomer conformation and the dimer organization, significant lower thermal stability than the mutant enzyme A143Q. The ratio of turnover number to Km-value is 1273fold lower than that for mutant enzyme A143Q 653320 3.4.21.97 A143Q/S225Y/L229R mutation has little effect on the stability of the dimer, mutant enzyme with large conformational changes with 40% lower helical content relative to the mutant enzyme A143Q, significant lower thermal stability than the mutant enzyme A143Q. The ratio of turnover number to Km-value is 1680fold lower than that for mutant enzyme A143Q 653320 3.4.21.97 A143T/A144T mutant displays better stability than wild-type and has esterase activity toward specific small ester compounds, e.g., Boc-L-Ala-4-nitrophenol 732540 3.4.21.97 A143V inability of the mutant virus to effect I-site cleavage in infected cells, mutation has no gross effect on the rate of virus production or on the amounts of extracellular virions, noninfectious enveloped particles and dense bodies 653074 3.4.21.97 H47A inactive mutant, active if coexpressed with AW5 which encodes the first 179 amino acids of assemblin with the addition of Ile, Gln, Thr 95597 3.4.21.97 H47A mutation prevents I-site cleavage (cleavage at an internal site of assemblin, converting it to a two-chain form that remain active). Imidazole restores I-site cleavage to mutant 665654 3.4.21.97 H61A mutation impairs protease activity. Morphology of mutant capsids exclusively resembles those of procapsids or large cored B capsids. Specifically, mutant capsids are spherical and lack electron dense cores. Most of these particles are tightly clustered in the nucleoplasm. Mutant does not replicate to appreciable extents on CV1 cells 732971 3.4.21.97 additional information IC-assemblin and IC-assemblinHis mutants of assemblin, blocked for internal and cryptic site cleavage. Enzymatic activities of pPR mutants are indistinguishable from that of IC-assemblin 681686 3.4.21.97 additional information stabilized against self-cleavage by a single point mutation in its cleavage site (IC-assemblin) 718057 3.4.21.97 S118A inactive mutant, active if coexpressed with AW5 which encodes the first 179 amino acids of assemblin with the addition of Ile, Gln, Thr 95597 3.4.21.97 S132A active-site mutant 95603 3.4.21.97 S132A is inactive 677592 3.4.21.97 S132A mutant enzyme is catalytically inactive 650245 3.4.21.97 S134A/A143Q the ratio of turnover number to Km-value is 150fold lower than that of the wild-typelike mutant enzyme A143Q 650071 3.4.21.97 S134A/A143Q/H157A the ratio of turnover number to Km-value is 174fold lower than that of the wild-typelike mutant enzyme A143Q 650071 3.4.21.97 V141G/V207G soluble and stable, indistinguishable from wild-type in vitro 95604