Abstract
A rapid double mixing technique has been applied to the reaction kinetics of enzymes usually analyzed by linked assay. The test enzyme is allowed to perform in the absence of the linking enzyme but then the convenience of enzymatic analysis of accumulated produce is exploited when a second mixing adds the linking enzyme. The analytical method is presented which enables the determination of the rate of product accumulation during the delay between mixings. Various advantages of the system in both steady-state and transient kinetic analyses are described. As an example, 3-phosphoglycerate kinase is assayed. The results address the proposal that the kinase interacts with the usual linking enzyme, glyceraldehyde-3-phosphate dehydrogenase, and also raise questions about product inhibition.
| Original language | English |
|---|---|
| Pages (from-to) | 444-448 |
| Number of pages | 5 |
| Journal | Analytical Biochemistry |
| Volume | 176 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 1 1989 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
Double mixing technique for linked enzyme assays. / Landgraf, Ralf; Betts, Graham F.
In: Analytical Biochemistry, Vol. 176, No. 2, 01.02.1989, p. 444-448.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Double mixing technique for linked enzyme assays
AU - Landgraf, Ralf
AU - Betts, Graham F.
PY - 1989/2/1
Y1 - 1989/2/1
N2 - A rapid double mixing technique has been applied to the reaction kinetics of enzymes usually analyzed by linked assay. The test enzyme is allowed to perform in the absence of the linking enzyme but then the convenience of enzymatic analysis of accumulated produce is exploited when a second mixing adds the linking enzyme. The analytical method is presented which enables the determination of the rate of product accumulation during the delay between mixings. Various advantages of the system in both steady-state and transient kinetic analyses are described. As an example, 3-phosphoglycerate kinase is assayed. The results address the proposal that the kinase interacts with the usual linking enzyme, glyceraldehyde-3-phosphate dehydrogenase, and also raise questions about product inhibition.
AB - A rapid double mixing technique has been applied to the reaction kinetics of enzymes usually analyzed by linked assay. The test enzyme is allowed to perform in the absence of the linking enzyme but then the convenience of enzymatic analysis of accumulated produce is exploited when a second mixing adds the linking enzyme. The analytical method is presented which enables the determination of the rate of product accumulation during the delay between mixings. Various advantages of the system in both steady-state and transient kinetic analyses are described. As an example, 3-phosphoglycerate kinase is assayed. The results address the proposal that the kinase interacts with the usual linking enzyme, glyceraldehyde-3-phosphate dehydrogenase, and also raise questions about product inhibition.
UR - http://www.scopus.com/inward/record.url?scp=0024546702&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0024546702&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(89)90340-0
DO - 10.1016/0003-2697(89)90340-0
M3 - Article
C2 - 2742135
AN - SCOPUS:0024546702
VL - 176
SP - 444
EP - 448
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 2
ER -