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- ABOUT HYPERBOLIC REGRESSION ANALYSIS
- ------------------------------------
-
- HYPER.EXE, Version 1.00, 1992
- Copyright J S Easterby
-
-
- Hyper.exe
- ---------
-
- Hyper.exe is a programme for the analysis of enzyme-kinetic data.
- The velocity (v) of enzyme-catalysed reactions is hyperbolically related
- to the substrate concentration ([S]) through the Michaelis-Menten
- equation:
-
- v = Vmax.[S]/(Km + [S])
-
- where Vmax and Km are kinetic constants characteristic of the particular
- enzyme and substrate.
-
- These constants are estimated in experiments in which [S] is varied and
- v measured. They are usually extracted by some linear transformation of
- the data such as the Lineweaver-Burk or Hanes plots. These transformations
- are unsatisfactory as they can result in artificial weighting of the data
- leading to erroneous estimates of Vmax and Km.
-
- A better approach is to fit the data directly to the best possible
- hyperbola. This is achieved by Hyper.exe using non-linear regression
- analysis. Lineweaver-Burk, Hanes, Eadie-Hofstee and Parameter Space plots
- and the kinetic constants derived from them are also provided for
- comparison.
-
- The linear plot equations are as follows:
-
- Hanes Plot:
- [S]/v = Km/Vm + [S]/Vm
-
- Lineweaver-Burk Plot:
- 1/v = (Km/Vm).(1/[S]) + 1/Vm
-
-
- Eadie-Hofstee Plot:
- v = Vm - (v/[S]).Km
-
- Parameter Space Plot:
-
- For any pair of v and [S] values there is an infinite
- number of Vm and Km values which will satisfy the
- Michaelis-Menten equation. In the parameter space plot
- Vm and Km are treated as the variables. For each data pair
- a line is drawn connecting v on the ordinate axis and [S]
- on the abscissa. For n data pairs there will be n such
- lines and these will have n(n-1)/2 intersections.
- The v and [S] values at the intersections correspond
- respectively to Vm and Km values satisfying the
- Michaelis-Menten equation for the corresponding two pairs
- of s\v data. The best estimates of Vm and Km for the data
- set as a whole are taken as the medians of the points
- of intersection.
-
- The programme should be of use in undergraduate teaching for the
- demonstration of the effects of experimental error and analysis method on
- the evaluation of kinetic constants but will also prove of day-to-day use
- for the analysis of kinetic data in Life Science laboratories.
-
-
- Installation
- ------------
-
- To install Hyper, run Windows 3.x and from within Program Manager or
- File Manager run A:\setup.exe. You will be prompted for an appropriate
- directory into which to install the programme.
-
- Hyper will require approximately 400-500K of free memory to load and
- Windows 3 standard or enhanced mode to run.
-
-
- Automatic Running and Loading of Data Files
- -------------------------------------------
-
- Data files for Hyper have the file extension .hyp. Hyper may be started
- with a data file as a command-line option (eg. Hyper data.hyp or just
- hyper data). Alternatively, Hyper may be run with no command-line option
- and the data subsequently entered or loaded from disk.
-
- The first time Hyper is run it will modify Win.ini to create an
- association between itself and .hyp files. Thereafter, 'double-clicking'
- a .hyp file or running one from the File or Program Managers will
- automatically load Hyper.exe. For this option to work Hyper.exe must be
- in a directory on your Path.
-
-
- Data File Format
- ----------------
-
- Data files should be saved with the extension .hyp. They may be created
- using any text editor or word processor and saved in text format.
- Substrate concentrations and enzyme velocities should either alternate
- on a line separated by tabs, spaces or linefeeds, or should be contained
- in two adjacent columns ([S] first).
-
- Data may also be pasted from editors, word processors and spreadsheets
- using the clipboard. Data may also be entered directly through the
- editor contained in Hyper.exe and saved to disk.
-
- Five example data files are supplied. These are named perfect.hyp,
- data1.hyp, data2.hyp, data3.hyp and data4.hyp. The first contains a
- short set of perfect hyperbolic data. data1 and Data2 contain poorer,
- simulated data with uncertainties of 20% and 30% respectively on the
- velocity measurements.
-
-
- Printing Data, Results and Graphs
- ---------------------------------
-
- Data and results may be copied to the clipboard or printed directly
- to the default printer.
-
- Graphical output of the hyperbola or other plots may also be copied to
- the clipboard or printed directly to the printer.
- To obtain the maximum size of graphic on the clipboard,
- the graphic screen should be maximised before copying.
-
-
- Calculation Speed
- -----------------
-
- You are limited to 100 [S],v pairs in this version of Hyper.
- For large data sets, calculation speed can be increased by iconising or
- not showing graphs during calculations. This obviates the need for
- updating graphics screens as data are changed or recalculation carried out.
-
- The parameter space calculations are slow for large sets of data and
- therefore the calculation has its own option button. The parameter space
- calculations are carried out in the background and may be interrupted by
- pressing the button a second time or by pressing the Esc (escape) key.
-
-
- Weighting
- ---------
-
- Simple weighting may be applied to velocity data. This assumes that the
- variance of the velocity is proportional to its square. Weighting may be
- chosen as a menu option.