1H‐NMR study of neurotoxin B‐IV from the marine worm Cerebratulus lacteus Solution properties, sequence‐specific resonance assignments, secondary structure and global fold

Poul Erik HANSEN, William R. KEM, Allan L. BIEBER, Raymond S. NORTON

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Sequence‐specific resonance assignments are reported for the 500‐MHz 1H‐NMR spectrum of the 55‐residue neurotoxin B‐IV, isolated from the heteronemertine worm Cerebratulus lacteus. A range of two‐dimensional homonuclear correlated and NOE spectra was used in making these assignments, which include NH, CαH and CβH resonances, as well as most resonances from longer‐chain spin systems, with the exception of the ten Lys residues, where spectral overlap prevented complete, unambiguous assignments. The secondary structure of B‐IV was identified from the pattern of sequential (i, i+ 1) and medium range (i, i+ 2/3/4) NOE connectivities and the location of slowly exchanging backbone amide protons. Two helices are present, incorporating residues 13–26 and 33–49, and the C‐terminal five residues form a helix‐like structure. A type‐I reverse turn, involving residues 28–31 is present in a small loop linking the two major helices, and the N‐terminus appears to be unordered at 27°C, although it may adopt a more ordered conformation at lower temperatures. These elements of secondary structure, together with the four disulfide bonds in the protein, provide sufficient information to define the global fold of the molecule in solution. The pH and temperature dependence of the toxin have been investigated by 1H‐NMR and the pKa values of several ionisable sidechains determined.

Original languageEnglish
Pages (from-to)231-240
Number of pages10
JournalEuropean Journal of Biochemistry
Issue number1
Publication statusPublished - 1992
Externally publishedYes

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