| Id |
Subject |
Object |
Predicate |
Lexical cue |
| T1 |
0-69 |
Sentence |
denotes |
Affinity labeling of spinach leaf phosphoribulokinase by ATP analogs. |
| T2 |
70-108 |
Sentence |
denotes |
Modification of an active site lysine. |
| T3 |
109-218 |
Sentence |
denotes |
Spinach leaf phosphoribulokinase is sensitive to modification by ATP analogs that react with lysine residues. |
| T4 |
219-318 |
Sentence |
denotes |
The 2',3'-dialdehyde derivative of ATP (oATP) inactivates enzyme in a slow, time-dependent fashion. |
| T5 |
319-484 |
Sentence |
denotes |
The process follows first-order kinetics (kinact = 0.07 min-1), and the concentration dependence of inactivation indicates tight inhibitor binding (Ki = 106 microM). |
| T6 |
485-610 |
Sentence |
denotes |
ATP offers good protection against inactivation (Kd = 67 microM), suggesting that oATP is directed toward the catalytic site. |
| T7 |
611-713 |
Sentence |
denotes |
This conclusion is supported by the fact that oATP functions as an alternate substrate (Km = 0.55 mM). |
| T8 |
714-815 |
Sentence |
denotes |
Inactivation of phosphoribulokinase by [14C]oATP results in a modification stoichiometry of 0.7/site. |
| T9 |
816-960 |
Sentence |
denotes |
The 14C-labeled enzyme is stable to dialysis, suggesting that the covalent adduct formed between protein and oATP is not a simple Schiff's base. |
| T10 |
961-1109 |
Sentence |
denotes |
Adenosine di- and triphosphopyridoxals (Ado-P2-Pl and Ado-P3-Pl, respectively) also inhibit spinach phosphoribulokinase in a time-dependent fashion. |
| T11 |
1110-1299 |
Sentence |
denotes |
In this case, activity loss is reversible unless the inhibited species is borohydride-reduced, suggesting that Ado-P2-Pl and Ado-P3-Pl form Schiff's bases with an amino group on the enzyme. |
| T12 |
1300-1398 |
Sentence |
denotes |
Protection is afforded by the substrate ATP, suggesting that modification is active site-directed. |
| T13 |
1399-1516 |
Sentence |
denotes |
Prolonged incubation of enzyme with these inhibitors does not result in complete inactivation of phosphoribulokinase. |
| T14 |
1517-1708 |
Sentence |
denotes |
Residual activity is dependent on inhibitor concentration, as would be expected if equilibrium is established between the noncovalent E.I complex and the covalent (Schiff's base) E-I species. |
| T15 |
1709-1814 |
Sentence |
denotes |
Kinetic data analysis indicates Ki values of 175 and 11 microM for Ado-P2-Pl and Ado-P3-Pl, respectively. |
| T16 |
1815-1933 |
Sentence |
denotes |
Thus, the ATP-binding domain can easily accommodate the pyridoxal moiety which is tethered to the polyphosphate chain. |
| T17 |
1934-2217 |
Sentence |
denotes |
The phosphorylated ATP analogs employed in this study exhibit substantially tighter binding to phosphoribulokinase than does fluorosulfonyl-benzoyladenosine (Ki = 4.8 mM), which we have previously demonstrated to be useful in selectively modifying the ATP-binding domain (Krieger, T. |
| T18 |
2218-2238 |
Sentence |
denotes |
J., and Miziorko, H. |
| T19 |
2239-2288 |
Sentence |
denotes |
M. (1986) Biochemistry 25, 3496-3501; Krieger, T. |
| T20 |
2289-2310 |
Sentence |
denotes |
J., Mende-Mueller, L. |
| T21 |
2311-2331 |
Sentence |
denotes |
M., and Miziorko, H. |
| T22 |
2332-2350 |
Sentence |
denotes |
M. (1987) Biochim. |
| T23 |
2351-2359 |
Sentence |
denotes |
Biophys. |
| T24 |
2360-2379 |
Sentence |
denotes |
Acta 915, 112-119). |
| T25 |
2380-2630 |
Sentence |
denotes |
Although the adduct formed between oATP and enzyme was unsuitable for structural analysis, borohydride reduction of the Schiff's base formed between enzyme and Ado-P3-[3H]Pl produced a species useful for investigation by protein chemistry techniques. |
| T26 |
2631-2775 |
Sentence |
denotes |
A radiolabeled tryptic peptide was prepared, isolated, and sequenced; the data indicate that lysine 68 is the residue modified by Ado-P3-[3H]Pl. |
| T1 |
0-69 |
Sentence |
denotes |
Affinity labeling of spinach leaf phosphoribulokinase by ATP analogs. |
| T2 |
70-108 |
Sentence |
denotes |
Modification of an active site lysine. |
| T3 |
109-218 |
Sentence |
denotes |
Spinach leaf phosphoribulokinase is sensitive to modification by ATP analogs that react with lysine residues. |
| T4 |
219-318 |
Sentence |
denotes |
The 2',3'-dialdehyde derivative of ATP (oATP) inactivates enzyme in a slow, time-dependent fashion. |
| T5 |
319-484 |
Sentence |
denotes |
The process follows first-order kinetics (kinact = 0.07 min-1), and the concentration dependence of inactivation indicates tight inhibitor binding (Ki = 106 microM). |
| T6 |
485-610 |
Sentence |
denotes |
ATP offers good protection against inactivation (Kd = 67 microM), suggesting that oATP is directed toward the catalytic site. |
| T7 |
611-713 |
Sentence |
denotes |
This conclusion is supported by the fact that oATP functions as an alternate substrate (Km = 0.55 mM). |
| T8 |
714-815 |
Sentence |
denotes |
Inactivation of phosphoribulokinase by [14C]oATP results in a modification stoichiometry of 0.7/site. |
| T9 |
816-960 |
Sentence |
denotes |
The 14C-labeled enzyme is stable to dialysis, suggesting that the covalent adduct formed between protein and oATP is not a simple Schiff's base. |
| T10 |
961-1109 |
Sentence |
denotes |
Adenosine di- and triphosphopyridoxals (Ado-P2-Pl and Ado-P3-Pl, respectively) also inhibit spinach phosphoribulokinase in a time-dependent fashion. |
| T11 |
1110-1299 |
Sentence |
denotes |
In this case, activity loss is reversible unless the inhibited species is borohydride-reduced, suggesting that Ado-P2-Pl and Ado-P3-Pl form Schiff's bases with an amino group on the enzyme. |
| T12 |
1300-1398 |
Sentence |
denotes |
Protection is afforded by the substrate ATP, suggesting that modification is active site-directed. |
| T13 |
1399-1516 |
Sentence |
denotes |
Prolonged incubation of enzyme with these inhibitors does not result in complete inactivation of phosphoribulokinase. |
| T14 |
1517-1708 |
Sentence |
denotes |
Residual activity is dependent on inhibitor concentration, as would be expected if equilibrium is established between the noncovalent E.I complex and the covalent (Schiff's base) E-I species. |
| T15 |
1709-1814 |
Sentence |
denotes |
Kinetic data analysis indicates Ki values of 175 and 11 microM for Ado-P2-Pl and Ado-P3-Pl, respectively. |
| T16 |
1815-1933 |
Sentence |
denotes |
Thus, the ATP-binding domain can easily accommodate the pyridoxal moiety which is tethered to the polyphosphate chain. |
| T17 |
1934-2217 |
Sentence |
denotes |
The phosphorylated ATP analogs employed in this study exhibit substantially tighter binding to phosphoribulokinase than does fluorosulfonyl-benzoyladenosine (Ki = 4.8 mM), which we have previously demonstrated to be useful in selectively modifying the ATP-binding domain (Krieger, T. |
| T18 |
2218-2238 |
Sentence |
denotes |
J., and Miziorko, H. |
| T19 |
2239-2288 |
Sentence |
denotes |
M. (1986) Biochemistry 25, 3496-3501; Krieger, T. |
| T20 |
2289-2310 |
Sentence |
denotes |
J., Mende-Mueller, L. |
| T21 |
2311-2331 |
Sentence |
denotes |
M., and Miziorko, H. |
| T22 |
2332-2350 |
Sentence |
denotes |
M. (1987) Biochim. |
| T23 |
2351-2359 |
Sentence |
denotes |
Biophys. |
| T24 |
2360-2379 |
Sentence |
denotes |
Acta 915, 112-119). |
| T25 |
2380-2630 |
Sentence |
denotes |
Although the adduct formed between oATP and enzyme was unsuitable for structural analysis, borohydride reduction of the Schiff's base formed between enzyme and Ado-P3-[3H]Pl produced a species useful for investigation by protein chemistry techniques. |
| T26 |
2631-2775 |
Sentence |
denotes |
A radiolabeled tryptic peptide was prepared, isolated, and sequenced; the data indicate that lysine 68 is the residue modified by Ado-P3-[3H]Pl. |