Abstract
Abstract A simple, rapid, manual technique is described for determining the amino-terminal amino acid sequence of proteins on a nanomole scale. In this modification of the 5-dimethylaminonaphthalene-1-sulfonyl-Edman degradation, inorganic carriers permit convenient manipulation of small amounts of protein, and use of the detergent sodium dodecyl sulfate throughout the procedure maintains protein solubility. Nanomole quantities of pure protein for such sequence analysis are readily isolated from multicomponent systems by analytical scale polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Proteins are recovered quantitatively from the gel by elution. The method is therefore suitable for characterization of the proteins derived from multichain enzymes and viruses.
Highlights
A simple, rapid, manual technique is described for determining the amino-terminal amino acid sequence of proteins on a nanomole scale
We have developed a reliable manual dansyl-Edman procedure which can be used to determine an amino-terminal sequence of 5 to 10 residues for any protein with an unblocked amino terminus which can be purified on a nanomole scale
More was used in several instances when sufficient starting material was available
Summary
A simple, rapid, manual technique is described for determining the amino-terminal amino acid sequence of proteins on a nanomole scale. The decision to determine a protein’s amino-terminal sequence depends on availability of the purified species in sufficient quantity Techniques such as the recently automated Edman degradation (1) have been extensively developed for proteins which can be purified in milligram amounts. We have developed a reliable manual dansyl-Edman procedure which can be used to determine an amino-terminal sequence of 5 to 10 residues for any protein with an unblocked amino terminus which can be purified on a nanomole scale. We have successfully applied our SDS-dansyl-Edman procedure to nanomole quantities of protein separated 011 and quantitatively eluted from SDS polyacrylamide gels. Proteins of both known and unknown amino-terminal sequence have been subjected to the SDS-dansyl-Edman degradation. For proteins of unknown sequence, such as minor capsid proteins from the coliphages Qfi and R17, our amino-terminal sequence data are compatible with known RNA sequences from the phage genome
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
