Mapping protein-protein interaction by 13C′-detected heteronuclear NMR spectroscopy

被引：24

作者：

Bertini, Ivano ^{[1
]}

Felli, Isabella C.

Gonnelli, Leonardo

Pierattelli, Roberta

Spyranti, Zinovia

Spyroulias, Georgios A.

机构：

[1] Univ Florence, Dept Chem, I-50019 Sesto Fiorentino, Italy

[2] Univ Patras, Dept Pharm, Patras 26504, Greece

[3] Univ Florence, CERM, I-50019 Sesto Fiorentino, Italy

来源：

JOURNAL OF BIOMOLECULAR NMR | 2006年 / 36卷 / 02期

关键词：

C-13-NMR; chemical shift perturbation; copper chaperones; metal-mediated interaction; proteins complex; protonless NMR;

D O I：

10.1007/s10858-006-9068-z

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The copper-mediated protein-protein interaction between yeast Atx1 and Ccc2 has been examined by protonless heteronuclear NMR and compared with the already available H-1-N-15 HSQC information. The observed chemical shift variations are analyzed with respect to the actual solution structure, available through intermolecular NOEs. The advantage of using the CON-IPAP spectrum with respect to the H-1-N-15 HSQC resides in the increased number of signals observed, including those of prolines. CBCACO-IPAP experiments allow us to focus on the interaction region and on side-chain carbonyls, while a newly designed CEN-IPAP experiment on side-chains of lysines. An attempt is made to rationalize the chemical shift variations on the basis of the structural data involving the interface between the proteins and the nearby regions. It is here proposed that protonless C-13 direct-detection NMR is a useful complement to H-1 based NMR spectroscopy for monitoring protein-protein and protein-ligand interactions.

引用

页码：111 / 122

页数：12