( the peptidic bonds adjacent to the cys...)Efficient Primary Structure Elucidation of Disulfide-bridged Peptides

the peptidic bonds adjacent to the cysteines, as illustrated in Figure 3. The identification of the disulfidebridge pattern of the CSTX peptides by tandem mass spectrometry represents a valuable alternative to traditional methods for elucidation of complex molecular structures. Modern highperformance instru- mentation, such as the QSTAR Pulsar mass spectrometer, greatly enhance the potential of this technique, since a wide range of information is obtained from a single experiment using minute amounts of sample only. Sample preparation is essentially reduced to a purification step, thus, avoiding the risk of potential structural alteration, e.g. disulfide scrambling.

Conclusions

CSTX-1 and CSTX-9 share the same disulfide bridge pattern linking Cys 1 Cys 4, Cys 2 Cys 5, Cys 3 Cys 8, Cys 6 Cys 7. This pattern is also found in other spider toxins, e.g. w-agatoxins and μ-agat oxins from Agelenopsis aperta, SNX-325 from Segestria florentina and curtatoxins from Hololena curta (Figure 3). This disulfide bridge pattern represents thewell-known consensus sequence of the inhibitor cystine knot structural motif: CX3-7CX3-10CX0-7CX1-8CX4-20C. The results demonstrate that Edman sequencing and tandem mass spectrometry are by no means competing techniques. These two techniques can be considered as complementary tools, each one offering its particular advantages. Edman sequencing is a straight forward and efficient approach to the determination of amino acid sequences of peptides and proteins, whereas tandem mass spectrometry exhibits its ultimate potential for the elucidation of structural details. If both techniques are available to the researcher, bioanalytical problems are solved most accurately and efficiently.

References

1L. Kuhn-Nentwig, J. Schaller, W. Nentwig, Purification of toxic peptides and the amino acid sequ
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