清华大学：《生物核磁共振波谱学》课程教学资源（PPT课件，英文版）6. 2D NMR Spectra

Welcome to the exiting world Ofmr though a little hard to learn remember Why just Mr How a ID MMR spectrum look like What we can obtain from it eand what can we do for a big molecule. THMMR

THNMR YAN Welcome to the exiting world of NMR… though a little hard to learn… remember… •Why just NMR … •How a 1D NMR spectrum look like… •What we can obtain from it… •and what can we do for a big molecule…

Information from ID NR Spectrum Integral Area Half Height Peak Width Amplitude CH CH J constants Peak splitting TMS NOE 5.0 3.0 2.0 00 T1 ppm T2 。 Chemiday8higen Exchange Chemical Shift Reference THMMR

Information from 1D NMR Spectrum THNMR YAN Chemical Shift Chemical Shift Reference J constants Peak Splitting Integral Area Amplitude Half Height Peak Width NOE T1 T2 … Hydrogen Exchange

/到 生物核磁共振波谱学 NMR in Biological Science 6. 2D NMR Spectra THNMR AN

生物核磁共振波谱学 NMR in Biological Science 6. 2D NMR Spectra THNMR YAN

6.1 NTRODUCTION 1971比利时 Jeener2DNMR思想 1974-76瑞士RR. Ernst2DNMR理论 1979~82瑞士K. Wuthrich2DNMR&蛋白质 1982~瑞士K. Wuthrich 2DNMR+距离几何一>蛋白质空间结构 荷兰 W.F. Van gunsteren&R. Kaptein 2DNMR+计算机分子动力学模拟一>空间结构 THNMR AN

THNMR YAN 6.1 INTRODUCTION 1971 比利时 Jeener 2D NMR思想 1974~76 瑞士 R.R. Ernst 2D NMR理论 1979~82 瑞士 K. Wuthrich 2D NMR & 蛋白质 1982~ 瑞士 K. Wuthrich 2D NMR +距离几何—> 蛋白质空间结构 荷兰W.F. Van Gunsteren & R. Kaptein 2D NMR + 计算机分子动力学模拟—> 空间结构

What is 2D NMR? FT 十a +t =nAt, FT FID信号的强度和相位与t2=0时 刻之前核自旋系统的状态有关 Ar+d s(tr, t2)--2D FT->SO1,O,) Pulse Sequence(Spin Echo Sequence) THMMR

THNMR YAN t1= nDt1 t2 What is 2D NMR? FID信号的强度和相位与t2=0时 刻之前核自旋系统的状态有关 S(t1 , t2 ) —2D FT—> S(w1 , w2 ) => Pulse Sequence (Spin Echo Sequence)

Scheme for 2D pulse sequences and experiments Preparation Evolution Mixing Acquisition t The first perturbation of the system(pulse)was called the preparation of the spin system The variable te is called the evolution time, tr We have a mixing event, in which information from one part of the spin system is relayed to other parts. Finally, we have an acquisition period (t2)as with all ID experiments THMMR

Scheme for 2D pulse sequences and experiments Preparation Evolution Acquisition t1 t2 Mixing THNMR YAN • The first perturbation of the system (pulse) was called the preparation of the spin system. • The variable tE is called the evolution time, t1 . • We have a mixing event, in which information from one part of the spin system is relayed to other parts. • Finally, we have an acquisition period (t2 ) as with all 1D experiments

What is 2D NMR? Preparation Evolution Magnetization Detection t2 FT(2) FT(11) S(142) 2 以异核13C-相关实验为例来说明2DNM实验的四个时期: 准备期( Preparation)、发展期( Evolution) 混合期( Magnetization transfer)和探测期( detection)

以异核13C－1H相关实验为例来说明2D NMR实验的四个时期： 准备期(Preparation)、发展期(Evolution)、 混合期(Magnetization transfer)和探测期(detection)。 What is 2D NMR? THNMR YAN

What is 2D NMR? √ 2D IR Yan YB, Luo XC, Zhou HM, Zhang RQ Chinese sci bull. 2002. 47: 389-393 THMMR Fabian H, Mantsch HH, Schultz CP. PNAS. 1999, 96: 13153-1315

THNMR YAN What is 2D NMR? Fabian H, Mantsch HH, Schultz CP. PNAS. 1999, 96: 13153–1315 Yan YB, Luo XC, Zhou HM, Zhang RQ. Chinese Sci Bull. 2002, 47: 389-393 2D IR

Typical pulse sequences used in(a) NMr spectroscopy and(b) heterodyned 2D-IR spectroscopy. t2 t3 The three pulses in both techniques are Pulse FID separated by time delays t1 and t 2, and the FID is measured versus t3 In heterodyned 几几几 2D-IR. the three ir laser beams and the emitted FID signal beam all travel in different directions, and thus are spatially separated from one another the electric field of the signal beam cannot be measured directly, as it is in NMR, so it is overlapped with a fourth local-oscillator beam that heterodynes the Current Opinion in Structural Biolog signal (not shown). When t2=0, the 2D-IR spectrum is analogous to a CosY experiment otherwise it is similar to noesy THMMR anni MT and Hochstrasser RM. Current Opinion in Structural Biology 2001, 11: 516-522

THNMR YAN The three pulses in both techniques are separated by time delays t1 and t2, and the FID is measured versus t3. In heterodyned 2D-IR, the three IR laser beams and the emitted FID signal beam all travel in different directions, and thus are spatially separated from one another. The electric field of the signal beam cannot be measured directly, as it is in NMR, so it is overlapped with a fourth local-oscillator beam that heterodynes the signal (not shown). When t2 = 0, the 2D-IR spectrum is analogous to a COSY experiment, otherwise it is similar to NOESY. Zanni MT and Hochstrasser RM. Current Opinion in Structural Biology 2001, 11:516–522 Typical pulse sequences used in (a) NMR spectroscopy and (b) heterodyned 2D-IR spectroscopy

Scheme for a 2D MR spectrum Diagonal signals. results from contributions of the magnetization that has not been lo, changed by the mixing sequence A (equal frequency in both dimensions) Cross signals originate from nuclei that exchanged magnetization during the mixing time (frequencies of the first and second nucleus in each dimension B respectively). They indicate an interaction of these two nuclei THMMR

Scheme for a 2D NMR spectrum THNMR YAN Diagonal signals: results from contributions of the magnetization that has not been changed by the mixing sequence (equal frequency in both dimensions). Cross signals: originate from nuclei that exchanged magnetization during the mixing time (frequencies of the first and second nucleus in each dimension, respectively). They indicate an interaction of these two nuclei