PMID- 10318778
OWN - NLM
STAT- MEDLINE
DCOM- 19990617
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 20
DP  - 1999 May 14
TI  - Contulakin-G, an O-glycosylated invertebrate neurotensin.
PG  - 13752-9
AB  - We have purified contulakin-G, a 16-amino acid O-linked glycopeptide
      (pGlu-Ser-Glu-Glu-Gly-Gly-Ser-Asn-Ala-Thr-Lys-Lys-Pro-Tyr-Ile-Leu-OH, pGlu is
      pyroglutamate) from Conus geographus venom. The major glycosylated form of
      contulakin-G was found to incorporate the disaccharide
      beta-D-Galp-(1-->3)-alpha-D-GalpNAc-(1-->) attached to Thr10. The C-terminal
      sequence of contulakin-G shows a high degree of similarity to the neurotensin
      family of peptides. Synthetic peptide replicates of Gal(beta-->3)
      GalNAc(alpha-->)Thr10 contulakin-G and its nonglycosylated analog were prepared
      using an Fmoc (9-fluorenylmethoxycarbonyl) protected solid phase synthesis
      strategy. The synthetic glycosylated con- tulakin-G, when administered
      intracerebroventricular into mice, was found to result in motor
      control-associated dysfunction observed for the native peptide. Contulakin-G was 
      found to be active at 10-fold lower doses than the nonglycosylated Thr10
      contulakin-G analog. The binding affinities of contulakin-G and the
      nonglycosylated Thr10 contulakin-G for a number of neurotensin receptor types
      including the human neurotensin type 1 receptor (hNTR1), the rat neurotensin type
      1 and type 2 receptors, and the mouse neurotensin type 3 receptor were
      determined. The binding affinity of the nonglycosylated Thr10 contulakin-G was
      approximately an order of magnitude lower than that of neurotensin1-13 for all
      the receptor types tested. In contrast, the glycosylated form of contulakin-G
      exhibited significantly weaker binding affinity for all of the receptors tested. 
      However, both contulakin-G and nonglycosylated Thr10 contulakin-G were found to
      be potent agonists of rat neurotensin receptor type 1. Based on these results, we
      conclude that O-linked glycosylation appears to be a highly unusual strategy for 
      increasing the efficacy of toxins directed against neurotransmitter receptors.
FAU - Craig, A G
AU  - Craig AG
AD  - The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La
      Jolla, California 92037, USA.
FAU - Norberg, T
AU  - Norberg T
FAU - Griffin, D
AU  - Griffin D
FAU - Hoeger, C
AU  - Hoeger C
FAU - Akhtar, M
AU  - Akhtar M
FAU - Schmidt, K
AU  - Schmidt K
FAU - Low, W
AU  - Low W
FAU - Dykert, J
AU  - Dykert J
FAU - Richelson, E
AU  - Richelson E
FAU - Navarro, V
AU  - Navarro V
FAU - Mazella, J
AU  - Mazella J
FAU - Watkins, M
AU  - Watkins M
FAU - Hillyard, D
AU  - Hillyard D
FAU - Imperial, J
AU  - Imperial J
FAU - Cruz, L J
AU  - Cruz LJ
FAU - Olivera, B M
AU  - Olivera BM
LA  - eng
SI  - GENBANK/AF121108
GR  - GM 48677/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA, Complementary)
RN  - 0 (Glycoproteins)
RN  - 0 (Neuropeptides)
RN  - 0 (contulakin-G protein, Conus geographus)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Carbohydrate Sequence
MH  - Chromatography, High Pressure Liquid
MH  - DNA, Complementary/chemistry
MH  - Glycoproteins/chemistry/*isolation & purification
MH  - Glycosylation
MH  - Humans
MH  - Mice
MH  - Molecular Sequence Data
MH  - Mollusca/*chemistry
MH  - Neuropeptides/chemistry/*isolation & purification
MH  - Protein Processing, Post-Translational
MH  - Rats
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
EDAT- 1999/05/13 00:00
MHDA- 1999/05/13 00:01
CRDT- 1999/05/13 00:00
PHST- 1999/05/13 00:00 [pubmed]
PHST- 1999/05/13 00:01 [medline]
PHST- 1999/05/13 00:00 [entrez]
AID - 10.1074/jbc.274.20.13752 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 May 14;274(20):13752-9. doi: 10.1074/jbc.274.20.13752.