The first gamma-carboxyglutamic acid-containing contryphan - A selective L-type calcium ion channel blocker isolated from the venom of conus marmoreus
(2004) In Journal of Biological Chemistry 279(31). p.32453-32463- Abstract
- Contryphans constitute a group of conopeptides that are known to contain an unusual density of post-translational modifications including tryptophan bromination, amidation of the C-terminal residue, leucine, and tryptophan isomerization, and proline hydroxylation. Here we report the identification and characterization of a new member of this family, glacontryphan-M from the venom of Conus marmoreus. This is the first known example of a contryphan peptide carrying glutamyl residues that have been post-translationally carboxylated to {gamma}-carboxyglutamyl (Gla) residues. The amino acid sequence of glacontryphan-M was determined using automated Edman degradation and electrospray ionization mass spectrometry. The amino acid sequence of the... (More)
- Contryphans constitute a group of conopeptides that are known to contain an unusual density of post-translational modifications including tryptophan bromination, amidation of the C-terminal residue, leucine, and tryptophan isomerization, and proline hydroxylation. Here we report the identification and characterization of a new member of this family, glacontryphan-M from the venom of Conus marmoreus. This is the first known example of a contryphan peptide carrying glutamyl residues that have been post-translationally carboxylated to {gamma}-carboxyglutamyl (Gla) residues. The amino acid sequence of glacontryphan-M was determined using automated Edman degradation and electrospray ionization mass spectrometry. The amino acid sequence of the peptide is: Asn-Gla-Ser-Gla-Cys-Pro-D-Trp-His-Pro-Trp-Cys. As with most other contryphans, glacontryphan-M is amidated at the C terminus and maintains the five-residue intercysteine loop. The occurrence of a D-tryptophan residue was confirmed by chemical synthesis and HPLC elution profiles. Using fluorescence spectroscopy we demonstrated that the Gla-containing peptide binds calcium with a KD of 0.63 mM. Cloning of the full-length cDNA encoding glacontryphan-M revealed that the primary translation product carries an N-terminal signal/propeptide sequence that is homologous to earlier reported contryphan signal/propeptide sequences up to 10 amino acids preceding the toxin region. Electrophysiological experiments, carried out on mouse pancreatic B-cells, showed that glacontryphan-M blocks L-type voltage-gated calcium ion channel activity in a calcium-dependent manner. Glacontryphan-M is the first contryphan reported to modulate the activity of L-type calcium ion channels. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/123312
- author
- Hansson, Karin M LU ; Ma, Xiaosong LU ; Eliasson, Lena LU ; Czerwiec, Eva ; Furie, Bruce ; Furie, Barbara C. ; Rorsman, Patrik LU and Stenflo, Johan LU
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 279
- issue
- 31
- pages
- 32453 - 32463
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- scopus:2942529410
- wos:000222849700059
- pmid:15155730
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.M313825200
- language
- English
- LU publication?
- yes
- id
- 482f5817-65a9-4992-92aa-0051dcbe9b59 (old id 123312)
- date added to LUP
- 2016-04-01 11:34:42
- date last changed
- 2022-04-20 18:46:33
@article{482f5817-65a9-4992-92aa-0051dcbe9b59, abstract = {{Contryphans constitute a group of conopeptides that are known to contain an unusual density of post-translational modifications including tryptophan bromination, amidation of the C-terminal residue, leucine, and tryptophan isomerization, and proline hydroxylation. Here we report the identification and characterization of a new member of this family, glacontryphan-M from the venom of Conus marmoreus. This is the first known example of a contryphan peptide carrying glutamyl residues that have been post-translationally carboxylated to {gamma}-carboxyglutamyl (Gla) residues. The amino acid sequence of glacontryphan-M was determined using automated Edman degradation and electrospray ionization mass spectrometry. The amino acid sequence of the peptide is: Asn-Gla-Ser-Gla-Cys-Pro-D-Trp-His-Pro-Trp-Cys. As with most other contryphans, glacontryphan-M is amidated at the C terminus and maintains the five-residue intercysteine loop. The occurrence of a D-tryptophan residue was confirmed by chemical synthesis and HPLC elution profiles. Using fluorescence spectroscopy we demonstrated that the Gla-containing peptide binds calcium with a KD of 0.63 mM. Cloning of the full-length cDNA encoding glacontryphan-M revealed that the primary translation product carries an N-terminal signal/propeptide sequence that is homologous to earlier reported contryphan signal/propeptide sequences up to 10 amino acids preceding the toxin region. Electrophysiological experiments, carried out on mouse pancreatic B-cells, showed that glacontryphan-M blocks L-type voltage-gated calcium ion channel activity in a calcium-dependent manner. Glacontryphan-M is the first contryphan reported to modulate the activity of L-type calcium ion channels.}}, author = {{Hansson, Karin M and Ma, Xiaosong and Eliasson, Lena and Czerwiec, Eva and Furie, Bruce and Furie, Barbara C. and Rorsman, Patrik and Stenflo, Johan}}, issn = {{1083-351X}}, language = {{eng}}, number = {{31}}, pages = {{32453--32463}}, publisher = {{American Society for Biochemistry and Molecular Biology}}, series = {{Journal of Biological Chemistry}}, title = {{The first gamma-carboxyglutamic acid-containing contryphan - A selective L-type calcium ion channel blocker isolated from the venom of conus marmoreus}}, url = {{http://dx.doi.org/10.1074/jbc.M313825200}}, doi = {{10.1074/jbc.M313825200}}, volume = {{279}}, year = {{2004}}, }