Table of Contents
{1} BINDING STUDIES
{2} Big problems and issues...
{3} Binding Experiments covered by these lectures:
{4} Irreversible binding or Titration with a "pulled” reaction
{5} Why is the end-point not sharp?
{6} Why...
{7} In addition, [L] often is detected in measuring [EL]
{8} One binding site, one:one interaction:
{9} Also...
{10} Typical one-to-one binding experiments
{11} Two types of experiments...
{12} No experimental constraints…
{13} However, the + sign doesn’t make sense, i.e. x increases indefinitely!
{14} Prior to computers and fancy calculators,
{15} What if x is only on a relative scale?
{16} Terms become dimensionless
{17} WARNING...
{18} With experimental constraint that Lo>>Eo
{19} This equation:
{20} These equations allow graphical determination of Kd:
{21} This model, Y = Lo/(Kd + Lo)
{22} Multiple Equivalent, Independent Sites
{23} Solution in terms of things we know (Eo and Lo):
{24} Langmuir like derivation,
{25} With Y = [AL]/nEo measured as Signal/Signalmax
{26} Y and R are fundamentally different.
{27} Remember:
{28} Like the Langmuir isotherm, this equation linearizes several ways:
{29} Other Multi-site Models and Cooperativity
{30} Consider two such sites:
{31} General Form of 2-site Equation:
{32} General forms of other multisite models:
{33} All of these equations (1-site, 2-site, 3-site and 4-site) generate a family of OVERLAPPING curves.
{34} Positive and Negative Cooperativity
{35} Negative Cooperativity
{36} Measures of Cooperativity:
{37} 2) The Hill Plot Slope, h
{38} Hill coefficient...
{39} 3) Scatchard Plot (BOO-HISS) Maximum, Ymax
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Author: John E. Wampler
Email: wampler@bchiris.bmb.uga.edu
Home Page: http://bmbiris.bmb.uga.edu/wampler/8010
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