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.
Sweatt
J D
JD
Howard Hughes Medical Institute, Center for Neurobiology and Behavior, College of Physicians and Surgeons of Columbia University, New York, NY 10032.
Kennedy
T E
TE
Wager-Smith
K
K
Gawinowicz
M A
MA
Barzilai
A
A
Karl
K A
KA
Kandel
E R
ER
D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia; D002490:Central Nervous System / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D007256:Information Systems; D011506:Proteins / Q000032:analysis*; D012697:Serine Endopeptidases / Q000494:pharmacology
D011506:Proteins; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase