User:Paul Rothenberg/Notebook/Pauls Notebook/2013/05/24

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Reaction of OPD and H2O2 n MeOH with Mb/Tris Catalyst

  • Objectives:
  • 1. Continue analysis of OPD+H2O2 reaction with Mb in Tris catalyst catalyst at the same temperatures

  • Procedure:
  • 1. Followed same procedure as 5/20 with Mb lyopholized in Tris/Citrate and KCl from 5/22

  • Observations:

The Mb in Tris had no anomaly peaks at any temperature, he Mb in Tris had the highest absorbances at 5C and 45C, and the Mb in Citrate did not produce results similar to the previous trials. There was no peak at either 420 nm or 460 nm, and the 45C trial had completely skewed data from the previous temperatures.


Mb Tris 5C Sequential Time Absorbance Graph.png

Mb Tris 15C WORKUP GRAPH.png

Mb Tris MeOH 15C Sequential Time Absorbance Graph.png

Mb Tris 25C WORKUP GRAPH.png

Mb Tris 25C Sequential Time Absorbance Graph.png

Mb Tris 35C WORKUP GRAPH.png

Mb Tris 35C Sequential Time Absorbance Graph.png

Mb Tris 45C WORKUP GRAPH.png

Mb Tris 45C Sequential Time Absorbance Graph.png

Reanalysis of Reaction of OPD and H2O2 with Mb/Citrate Catalyst in MeOH

Mb Citrate 5C SEQUENTIAL May 24 WORKUP.png

Mb Citrate 15C May 24 WORKUP GRAPH.png

Mb Citrate 25C May 24 WORKUP GRAPH.png

Mb Citrate 45C May 24 WORKUP GRAPH.png

Mb Citrate May 24 350NM Time Absorbance GRAPH.png

Mb Citrate May 24 420NM Time Absorbance GRAPH.png

From this data, it seemed there may be some reaction between citrate and myoglobin directly. Some research was done into this, and there could be some correlation leading to the inconsistent data. This article was interesting in the relationship between myoglobin and citrate. Here is an article about low pH Myoglobin photoproducts.

Just found another article about metastable intermediates in myoglobin at low pH. Link is here. The authors discuss that at a low pH, the proximal histidine is protonated, allowing for the uptake of, as they mention, CO, or any other compound. CO may not necessarily be what's binding in this case. If we can determine what products are formed in the reaction, we can infer more about the reactivity of the overall protein and the iron center.

(added on 5/30).

  • References:

1. Lin et al, Myoglobin content and citrate synthase activity in different parts of the normal human heart, Journal of Applied Physiology, 1990, 69(3), 899-901. Found here.