Letters in Peptide Science

, Volume 4, Issue 1, pp 1–11 | Cite as

Identification of a binding region onEscherichia coli heat-stable enterotoxin to intestinal guanylyl cyclase C

  • Makoto Hasegawa
  • Yuki Kawano
  • Kazuya Matsumoto
  • Yuji Hidaka
  • Takashi Sato
  • Yasutsugu Shimonishi
Article

Summary

The heat-stable enterotoxin (ST), produced byEscherichia coli, causes acute diarrhea in infants and domestic animals by activation of the intestinal membrane-bound receptor, guanylyl cyclase C. We have investigated a region on the ST molecule, which is recognized by the receptor, by introducing a photochromophore,p-azidophenylalanine (Pap), into three different regions of STp(4–17), which has the full toxic activity. Each ST analog bound to the receptor, but only STp(4–17) containing a Pap residue at position 11 in the central portion of the ST molecule, showed a high efficiency in the reaction which cross-linked with the receptor by UV radiation. These data clearly demonstrate that the region of the ST molecule encompassing the Asn11 residue directly interacts with the receptor.

Keywords

Photoaffinity labeling Cross-linking p-Azidophenylalanine 

Abbreviations

ST

heat-stable enterotoxin produced by enterotoxigenicEscherichia coli

E. coli

Escherichia coli

STp

ST elaborated by a strain of enterotoxigenicE. coli isolated from pig

GC-C

guanylyl cyclase C

Aph

p-amino-l-phenylalanine

Pap

p-azido-l-phenylalanine

RP-HPLC

reversed-phase high-performance liquid chromatography

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis

CD

circular dichroism

ELISA

enzyme-linked immunosorbent assay

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Copyright information

© ESCOM Science Publishers 1997

Authors and Affiliations

  • Makoto Hasegawa
    • 1
  • Yuki Kawano
    • 1
  • Kazuya Matsumoto
    • 1
  • Yuji Hidaka
    • 1
  • Takashi Sato
    • 1
  • Yasutsugu Shimonishi
    • 1
  1. 1.Division of Protein Organic Chemistry, Institute for Protein ResearchOsaka UniversitySuita-shi, OsakaJapan

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