Sequential assignment and structure determination of spider toxin ω-aga-IVB 1,2

Hongtao Yu, Michael K. Rosen, Nicholas A. Saccomano, Douglas Phillips, Robert A. Volkmann, Stuart L. Schreiber

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

The solution structure of a peptide toxin isolated from funnel web spider venom, ω-Aga-IVB, was determined by 2D NMR methods. ω-Aga-IVB is a high-affinity specific blocker of P-type voltagedependent calcium channels. Nearly all of the proton resonances of this 48-residue protein were assigned using conventional 2D homonuclear NMR experiments. The three-dimensional structure of the molecule was determined by simulated annealing. The distance and dihedral restraints used in the structure calculations were derived from NOESY and COSY-type experiments, respectively. Mass spectrometric analysis of ω-Aga-IVB suggests that the protein contains four isulfide bonds. In the absence of chemical data to identify the pattern of cysteine pairing, the disulfide bonds of the toxin are proposed from the NMR data and subsequent structural calculations. The structure of the toxin can be described as a three-stranded anti-parallel β sheet connected by flexible loops. A striking feature of the structure is that the C-terminal 10 residues of this protein adopt random coil conformations. Several positively charged amino acid side chains are found localized on one face of the molecule, in close proximity to the C-terminal tail. This observation has led us to propose a speculative model of the toxins blockade mechanism.

Original languageEnglish (US)
Pages (from-to)13123-13129
Number of pages7
JournalBiochemistry®
Volume32
Issue number48
StatePublished - 1993

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Spiders
Nuclear magnetic resonance
P-Type Calcium Channels
Proteins
Molecules
Simulated annealing
Disulfides
Cysteine
Conformations
Protons
Tail
Experiments
Amino Acids
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Yu, H., Rosen, M. K., Saccomano, N. A., Phillips, D., Volkmann, R. A., & Schreiber, S. L. (1993). Sequential assignment and structure determination of spider toxin ω-aga-IVB 1,2. Biochemistry®, 32(48), 13123-13129.

Sequential assignment and structure determination of spider toxin ω-aga-IVB 1,2. / Yu, Hongtao; Rosen, Michael K.; Saccomano, Nicholas A.; Phillips, Douglas; Volkmann, Robert A.; Schreiber, Stuart L.

In: Biochemistry®, Vol. 32, No. 48, 1993, p. 13123-13129.

Research output: Contribution to journalArticle

Yu, H, Rosen, MK, Saccomano, NA, Phillips, D, Volkmann, RA & Schreiber, SL 1993, 'Sequential assignment and structure determination of spider toxin ω-aga-IVB 1,2', Biochemistry®, vol. 32, no. 48, pp. 13123-13129.
Yu H, Rosen MK, Saccomano NA, Phillips D, Volkmann RA, Schreiber SL. Sequential assignment and structure determination of spider toxin ω-aga-IVB 1,2. Biochemistry®. 1993;32(48):13123-13129.
Yu, Hongtao ; Rosen, Michael K. ; Saccomano, Nicholas A. ; Phillips, Douglas ; Volkmann, Robert A. ; Schreiber, Stuart L. / Sequential assignment and structure determination of spider toxin ω-aga-IVB 1,2. In: Biochemistry®. 1993 ; Vol. 32, No. 48. pp. 13123-13129.
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AU - Schreiber, Stuart L.

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