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

FEBS Letters

Iijima, H.¹  Amodeo, P.³  Temussi, P.A.³  Tancredi, T.²  Saviano, G.³ 

[1] Kirin Research Laboratories, Japan;ICMIB, CNR, via Tolano, Arco Felice, Napoli, Italy;Dipartimento di Chimica Università di Napoli Federico II, via Mezzocannone 4, Napoli, Italy

关键词: Sweet receptor;    Monellin;    NMR;    Molecular mechanics;    pMNEI;    single chain monellin;    NMR;    nuclear magnetic resonance;    NOE;    nuclear Overhauser effect;    DQF-COSY;    double-quantum filtered correlation spectroscopy;    TOCSY;    total correlation spectroscopy;    NOESY;    nuclear Overhauser effect spectroscopy;    MCD;    main chain directed assignment;    MM;    molecular mechanics;   

DOI  :  10.1016/0014-5793(92)81138-C

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 β-sheet of pMNEI that contains a likely ‘sweet finger’: the loop connecting the β-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 or 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 or aspartame.

【 授权许可】

Unknown