A test case of correlation metric construction of a reaction pathway from measurements

Science

Volumn 277, Issue 5330, 1997, Pages 1275-1279

  Arkin, Adam ^a   Shen, Peidong ^a   Ross, John ^a  

^a STANFORD UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ADENOSINE PHOSPHATE; CITRIC ACID; DIHYDROXYACETONE PHOSPHATE; ENZYME; GLUCOSE; GLYCERALDEHYDE 3 PHOSPHATE;

ALGORITHM; ARTICLE; CAPILLARY ZONE ELECTROPHORESIS; CHEMICAL REACTION KINETICS; GLYCOLYSIS; METRIC SYSTEM; PRIORITY JOURNAL; REACTION ANALYSIS; REACTOR; THEORY;

EID: 0030848624     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5330.1275     Document Type: Article

References (18)

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5. Solution was flowed into the CSTR through a peristaltic pump at a rate of 0.2 ml/min. A 10,000-MW ultrafiltration membrane (YM 10 Diaflo by Amicon, Beverly, MA) was used to confine the enzymes to the reactor. The solution is stirred with a Teflon stirring disk, which reduced the reactor volume to 1.17 ml.
6. P. J. Oefner, Electrophoresis 16, 46 (1995).
7. i, inorganic phosphate; and creatine-P, creatine phosphate.
8. P. Shen, D. Hauri, J. Ross, P. J. Oefner, J. Capillary Electrophor. 3, 155 (1996).
9. All chemicals and enzymes were purchased from Sigma, except for PFK2::F26BPase, which was kindly provided by D. Okar. Reactions were carried out in a solution of 30 mM Hepes. 50 mM potassium chloride, 1.0 mM potassium phosphate, 2 mM dithiothreitol, 5.0 mM magnesium chloride, 0.8 mM adenosine triphosphate (ATP), 2.5 mM creatine-P, and 0.6 mM glucose, buffered at pH 7.1 with sodium hydroxide, which we will refer to as enzyme buffer. All enzymes are from rabbit muscle unless otherwise indicated. The enzymes include HK from baker's yeast; PHI, PFK1, and F16BP from rabbit liver; and aldolase, TPI, CK, and PFK2::F26BPase from rat liver.
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15. The enzyme activities were not constant during the entire 12-hour sampling period. Six separate experiments of 2 to 3 hours each were conducted, with the same amount of enzymes (by weight) in each experiment. Nonetheless, the enzyme activities differed slightly between experiments, as indicated by a change in the unperturbed steady-state concentrations of the measured species. In order to make the time series between experiments match, we multiplied the concentrations from the time series in each experiment by the appropriate ratio of the reference concentration (as defined above) to the measured unperturbed steady-state concentration. In order to repeat the last time point of a previous experiment, each experiment repeated the last two to three sets of input from the previous experiment.
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18. We thank P. Oefner (Department of Biochemistry, Stanford University) for the use of the capillary electrophoresis apparatus and for helpful discussions, and S. Pilkis (deceased) and D. Okar (Department of Biochemistry, University of Minnesota) for providing us with PFK2::F26BPase and for helpful discussions. This work was supported in part by NSF. A.A. was supported in part by Office of Naval Research grant N00014-96-1-0564.