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Isolation of Low Molecular Weight Digestion Products of the Human Platelet Thromboxane A2 Receptor, Rev A

phoresis with the technique of tricine gel electrophoresis.

In this regard, the recent development of continuous elution preparative gel electrophoresis has provided significant benefits to the protein biochemist. The Mini Prep Cell enables fractionation and isolation of a mixture of proteins with minimal sample loss. In addition, complete isolation can be achieved within a shorter time frame, since commonly used electroelution techniques for the recovery of proteins from gel slabs are not needed. The development of tricine gel electrophoresis has also provided significant advantages in analytical protein chemistry, since it allows effective resolution of low molecular weight proteins at relatively moderate acrylamide concentrations. Thus, substitution of tricine for glycine as the trailing ion,7 results in low molecular weight resolution which cannot be achieved using the conventional glycine gel electrophoresis procedure of Laemmli.8 An added advantage of this approach is that the use of tricine versus glycine prevents signal disruption during subsequent amino acid sequencing. In light of these considerations, we have developed a procedure for casting tricine tube gels in the continuous elution preparative gel electrophoresis Mini Prep Cell. This procedure was then used to successfully isolate and accumulate low molecular weight digestion products of the human platelet TXA2 receptor.


Methods

SAMPLE PREPARATION

Purified TXA2 receptor protein was chemically digested with cyanogen bromide according to the method described by Gross.9 In short, receptor protein was suspended in 70% formic acid,
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