Development of a database of amino acid sequences for proteins identified and isolated on two‐dimensional polyacrylamide gels

As part of our continuing studies into the biochemical basis of long‐term changes in neuronal function in Aplysia, we have developed a simple method for obtaining amino acid sequence information from proteins isolated on two‐dimensional gels. Proteins isolated on preparative two‐dimensional gels are digested in situ with Staphylococcus, aureus V8 protease, and the resulting peptides electrophoresed, transferred to a polyvinylidene difluoride membrane, and sequenced in a gas‐phase sequencer. The method is simple and should be applicable to a variety of other systems where the development of a two‐dimensional gel database is underway.