Structural determination of the active site of a sweet protein. A 1H NMR investigation of pMNEI

FEBS Lett. 1992 Sep 21;310(1):27-30. doi: 10.1016/0014-5793(92)81138-c.

Abstract

pMNEI, a single chain sweet protein related to monellin, has been studied by means of 1H NMR at 500 MHz. A partial sequential assignment performed by means of the MCD method allowed the determination of the secondary structure of a large portion of the beta-sheet of pMNEI that contains a likely 'sweet finger': the loop connecting the beta-strands from residue 59 to residue 78, corresponding to segment 16-35 of the A chain of monellin. The detailed three-dimensional structure of the loop (Tyr66-Ala67-Ser68-Asp69), determined from several interresidue and intraresidue NOEs and subsequent energy minimization, shows that the side chains of Tyr66 and Asp69 fit our model of the sweet receptor in a manner very similar to that of the side chains of Phe and Asp of aspartame.

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Sequence Data
  • Plant Proteins / chemistry*
  • Protein Conformation
  • Sweetening Agents / chemistry*

Substances

  • Plant Proteins
  • Sweetening Agents
  • monellin protein, Dioscoreophyllum cumminsii