[Protein-analysis] Re: carbonic saturated distilled water?

Being Riptup brewhaha at freenet.edmonton.ab.ca
Thu Jul 20 02:40:51 EST 2006


I wrote this to ensure that Nina doesn't forget what she may have
learned about matters of choice when she does the write-up, and to
ensure that she doesn't misinterpret my speculation. I hav no idea
which tissues contain most of this enzyme, and I guess that's her
ultimate purpose. Wouldn't it be funky if the effect  of the enzyme was
actually to make carbonate (in the presence of zinc) _unstable_ on the
outer surface of lung tissue, so that inhibiting the enzyme would
result in carbonate retention?

==Begin Fiction===
Objective:   To assay the activity of carbonic anhydrase in bovine lung
tissue.

Hypothesis: No activity. The enzyme locates zinc in tissue where it
should
                  catalyze the formation of carbonate, on the outside
of bovine lung tissue.

Procedure:  (Sample may be too large for containment of exhaust).
                  (Ratio more important, and a large sample reduces
error).
                  Isolated carbonic anhydrase (structure: , purity: X%,
plus or minus) in a
                     solution of 1.1 L.
                  Assayed .1 L of the isolate for zinc content by
incineration and
                     precipitation.
                  Prepared a control or baseline medium of zinc
carbonate with the
                     same molar concentration of zinc and carbonate as
the enzyme solution.
                  Extracted CO2 with vacuum pressure.
                  Since unequal vacuum pressures will contaminate one
sample's
                      exhaust with more water, pressure will be
equalized.
                  Reversed reaction by applying exhaust gas with
positive pressure.
                  Curves of forward and backward reaction, in reverse
of time, _might_ match.

Results: Reduced pressure in the ionic solution to the point where it
expelled carbon dioxide at one mole per minute. At the same pressue,
the zinc-enzyme solution expelled X moles of carbon dioxide per minute.
Equalized pressure after T minutes. When I adjusted pressure in the
zinc-enzyme solution to equality with the control, the zinc-enzyme
solution expelled Y moles of carbon dioxide per minute. It is therefore
has an activity ratio in relation to the ionic solution of A/B moles
per minute.

              Arrived at hydrocarbonate stage where half of the CO2 has
been removed,
              so that bicarbonate dominates. The bicarbonate activity
level is C/D zinc-
              enzyme hydrocarbonate to zinc hydrocarbonate solution.
Continued
              experiment until some degree of near completion to allow
extrapolation of
              curve to completion.
              Applied exhaust gas to each solution at a pressure
                   resulting in carbonation at one mole per minute for
the ionic solution,
                   and at an equal pressure for the organic solution.
              Time required for organic solution to consume exhaust:
              Time required for ionic solution to consume exhaust:

Conclusions:
              Activity Ratio (mean or median):
                  Organic exhaust rate to ionic exhaust rate at start:
                  Organic exhaust rate to ionic exhaust rate at HCO3-
stage:
                  Organic carbonation rate to ionic rate at finish:
                  Organic carbonation rate to ionic rate at HCO3-
stage:
              (Optimally, four reaction rate curves,
              two plotted in reverse of time against the other two.)
              Estimate of error (percentage of maximum difference):
              Note that reaction in vivo doesn't need significant
pressure.
==End Fiction===
Doing this neatly (with the curves) might require some nifty equipment,
so maybe I should put it up at the halfbakery. :-)



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