Classification of amino acid spin systems using PFG HCC(CO)NH-TOCSY with constant-time aliphatic13C frequency labeling

✍ Scribed by Mitsuru Tashiro; Carlos B. Rios; Gaetano T. Montelione

• Publisher: Springer Netherlands
• Year: 1995
• Tongue: English
• Weight: 469 KB
• Volume: 6
• Category: Article
• ISSN: 0925-2738
• DOI: 10.1007/bf00211785

✦ Synopsis

We have developed a useful strategy for identifying amino acid spin systems and side-chain carbon resonance assignments in small 15N-, 13C-enriched proteins. Multidimensional constant-time pulsed field gradient (PFG) HCC(CO)NH-TOCSY experiments provide side-chain resonance frequency information and establish connectivities between sequential amino acid spin systems. In PFG HCC(CO)NH-TOCSY experiments recorded with a properly tuned constant-time period for frequency labeling of aliphatic 13C resonances, phases of cross peaks provide information that is useful for identifying spin system types. When combined with 13C chemical shift information, these patterns allow identification of the following spin system types: Gly, Ala, Thr, Val, Leu, Ile, Lys, Arg, Pro, long-type (i.e., Gln, Glu and Met), Ser, and AMX-type (i.e., Asp, Asn, Cys, His, Phe, Trp and Tyr).