3870137211

STRATEGIE NMR WYZNACZANIA STRUKTUR BLAŁEK W ROZTWORZE 25

ABSTRACT

A number of reasons have hindered the use of NMR spectroscopy as a tool for the protein structure determination. Recently, the ad\ ance in the NMR eąuipment, spectral techniques and isotope labelling has resulted in an enormous growth of NMR-determined protein structures.

After a brief presentation of protein structure and conformation several types of NMR-derived constraints and the characteristic features of Chemical shifts in prote-ins arc discusscd. Short-range, distancc and dihedral anglc constraints are \ aluable, but cumulativc crrors can appear when succcshc constraints are used to detcrmine spatial rclationship of remote parts of a protein Thcrcforc, long-rangc constraints dcrivcd from rcsidual dipolar couplings arc highh' complcmcntary to the short-range constraints.

Modem strategies to the NMR-based protein structure determination depend on the size of studied biomoleeules. Smali proteins (M_cz <10 kDa) can be studied with the use of two-dimensional (2D) NMR techniąues. Medium size proteins (M_cz< 30 kDa) reąuire double isotopic Iabeling ¹⁵N/¹³C and multidimensional (3D and 4D) heteronuclear techniąues. Thcre is no well established strategy to the structure determination of large proteins (M_cz >30 kDa) yet The most promising appro-aches take advantage of triple isotopic Iabeling ¹⁵N/¹³C/²H and the transverse relaxa-tion optimized spectroscopy (TROSY), both resulting in the reduction of signal width.

Kevwords: NMR spectrometry, protein structure in solution, NMR structural constrains

Słowa kluczowe: spektrometria NMR, struktury' białek w roztworach, więzy strukturalne NMR