[{"data":1,"prerenderedAt":1347},["ShallowReactive",2],{"mdc-sy2lni-key":3},{"data":4,"body":5},{},{"type":6,"children":7},"root",[8,17,24,31,37,42,46,52,57,62,87,92,95,101,106,117,122,140,143,149,154,172,175,181,187,192,197,202,215,218,224,229,234,252,257,270,273,279,284,289,302,307,320,323,329,334,337,343,348,353,366,371,374,380,385,462,467,470,476,481,510,515,518,524,529,535,553,556,562,568,580,585,590,595,598,604,615,620,625,630,633,639,649,654,659,662,668,674,679,685,690,696,701,739,744,747,753,758,763,776,781,786,791,794,800,805,810,815,818,824,847,850,856,861,866,871,874,880,885,903,908,913,918,921,927,932,937,940,946,952,957,963,968,973,978,983,986,992,997,1000,1006,1011,1016,1019,1025,1030,1035,1052,1057,1062,1067,1072,1077,1080,1086,1091,1095,1113,1118,1121,1127,1132,1155,1160,1165,1178,1183,1186,1192,1197,1240,1245,1248,1254,1267,1272,1275,1281,1286,1304,1309,1337,1342],{"type":9,"tag":10,"props":11,"children":13},"element","h1",{"id":12},"_1-why-is-protein-nmr-different-difficult-pages-15",[14],{"type":15,"value":16},"text","🧲 1. Why is Protein NMR Different & Difficult? (Pages 1–5)",{"type":9,"tag":18,"props":19,"children":21},"h2",{"id":20},"problem-i-signal-overlap",[22],{"type":15,"value":23},"🔹 Problem I: Signal Overlap",{"type":9,"tag":25,"props":26,"children":28},"h3",{"id":27},"page-2-single-amino-acid",[29],{"type":15,"value":30},"Page 2 – Single Amino Acid",{"type":9,"tag":32,"props":33,"children":34},"p",{},[35],{"type":15,"value":36},"A single amino acid → easy spectrum.\nEach proton (HN, Hα, Hβ, etc.) gives a separate peak.",{"type":9,"tag":32,"props":38,"children":39},{},[40],{"type":15,"value":41},"No ambiguity. No overlap.",{"type":9,"tag":43,"props":44,"children":45},"hr",{},[],{"type":9,"tag":25,"props":47,"children":49},{"id":48},"page-3-hexapeptide",[50],{"type":15,"value":51},"Page 3 – Hexapeptide",{"type":9,"tag":32,"props":53,"children":54},{},[55],{"type":15,"value":56},"Now peaks overlap heavily.",{"type":9,"tag":32,"props":58,"children":59},{},[60],{"type":15,"value":61},"Why?",{"type":9,"tag":63,"props":64,"children":65},"ul",{},[66,72,77,82],{"type":9,"tag":67,"props":68,"children":69},"li",{},[70],{"type":15,"value":71},"Many similar chemical environments",{"type":9,"tag":67,"props":73,"children":74},{},[75],{"type":15,"value":76},"Many NHs around 8 ppm",{"type":9,"tag":67,"props":78,"children":79},{},[80],{"type":15,"value":81},"Many aliphatic protons 0–4 ppm",{"type":9,"tag":67,"props":83,"children":84},{},[85],{"type":15,"value":86},"Aromatics pile up 6–8 ppm",{"type":9,"tag":32,"props":88,"children":89},{},[90],{"type":15,"value":91},"Result: impossible to interpret in 1D.",{"type":9,"tag":43,"props":93,"children":94},{},[],{"type":9,"tag":25,"props":96,"children":98},{"id":97},"page-4-148-aa-calmodulin",[99],{"type":15,"value":100},"Page 4 – 148 aa Calmodulin",{"type":9,"tag":32,"props":102,"children":103},{},[104],{"type":15,"value":105},"Massive 1D overlap.",{"type":9,"tag":32,"props":107,"children":108},{},[109,115],{"type":9,"tag":110,"props":111,"children":112},"strong",{},[113],{"type":15,"value":114},"Solution: Spread information into more dimensions.",{"type":15,"value":116},"\nInstead of 1D → use 2D and 3D.",{"type":9,"tag":32,"props":118,"children":119},{},[120],{"type":15,"value":121},"Think of it like:",{"type":9,"tag":63,"props":123,"children":124},{},[125,130,135],{"type":9,"tag":67,"props":126,"children":127},{},[128],{"type":15,"value":129},"1D = traffic jam",{"type":9,"tag":67,"props":131,"children":132},{},[133],{"type":15,"value":134},"2D = spread cars onto two roads",{"type":9,"tag":67,"props":136,"children":137},{},[138],{"type":15,"value":139},"3D = multi-level parking",{"type":9,"tag":43,"props":141,"children":142},{},[],{"type":9,"tag":25,"props":144,"children":146},{"id":145},"page-5-size-dependence",[147],{"type":15,"value":148},"Page 5 – Size Dependence",{"type":9,"tag":32,"props":150,"children":151},{},[152],{"type":15,"value":153},"HSQC spectra of proteins from 42 aa to 377 aa show:",{"type":9,"tag":63,"props":155,"children":156},{},[157,162,167],{"type":9,"tag":67,"props":158,"children":159},{},[160],{"type":15,"value":161},"More residues → more peaks",{"type":9,"tag":67,"props":163,"children":164},{},[165],{"type":15,"value":166},"Larger proteins → broader peaks (relaxation)",{"type":9,"tag":67,"props":168,"children":169},{},[170],{"type":15,"value":171},"Eventually impossible without isotope labeling",{"type":9,"tag":43,"props":173,"children":174},{},[],{"type":9,"tag":10,"props":176,"children":178},{"id":177},"_2-problem-ii-t-relaxation-pages-69",[179],{"type":15,"value":180},"🧲 2. Problem II: T₂ Relaxation (Pages 6–9)",{"type":9,"tag":25,"props":182,"children":184},{"id":183},"page-6-t-relaxation",[185],{"type":15,"value":186},"Page 6 – T₂ Relaxation",{"type":9,"tag":32,"props":188,"children":189},{},[190],{"type":15,"value":191},"Large proteins tumble slowly.",{"type":9,"tag":32,"props":193,"children":194},{},[195],{"type":15,"value":196},"Slow tumbling → short T₂ → broad peaks → low intensity.",{"type":9,"tag":32,"props":198,"children":199},{},[200],{"type":15,"value":201},"Key concept:",{"type":9,"tag":63,"props":203,"children":204},{},[205,210],{"type":9,"tag":67,"props":206,"children":207},{},[208],{"type":15,"value":209},"Small molecule → sharp peaks",{"type":9,"tag":67,"props":211,"children":212},{},[213],{"type":15,"value":214},"Large protein → broad weak peaks",{"type":9,"tag":43,"props":216,"children":217},{},[],{"type":9,"tag":25,"props":219,"children":221},{"id":220},"page-7-deuteration",[222],{"type":15,"value":223},"Page 7 – Deuteration",{"type":9,"tag":32,"props":225,"children":226},{},[227],{"type":15,"value":228},"Replace many ¹H with ²H (deuterium).",{"type":9,"tag":32,"props":230,"children":231},{},[232],{"type":15,"value":233},"Why it helps:",{"type":9,"tag":63,"props":235,"children":236},{},[237,242,247],{"type":9,"tag":67,"props":238,"children":239},{},[240],{"type":15,"value":241},"Fewer dipolar interactions",{"type":9,"tag":67,"props":243,"children":244},{},[245],{"type":15,"value":246},"Slower relaxation",{"type":9,"tag":67,"props":248,"children":249},{},[250],{"type":15,"value":251},"Narrower peaks",{"type":9,"tag":32,"props":253,"children":254},{},[255],{"type":15,"value":256},"Comparison shown:",{"type":9,"tag":63,"props":258,"children":259},{},[260,265],{"type":9,"tag":67,"props":261,"children":262},{},[263],{"type":15,"value":264},"Fully protonated → messy",{"type":9,"tag":67,"props":266,"children":267},{},[268],{"type":15,"value":269},"75% deuterated → clean",{"type":9,"tag":43,"props":271,"children":272},{},[],{"type":9,"tag":25,"props":274,"children":276},{"id":275},"page-8-trosy",[277],{"type":15,"value":278},"Page 8 – TROSY",{"type":9,"tag":32,"props":280,"children":281},{},[282],{"type":15,"value":283},"TROSY selects the slowest relaxing component of the multiplet.",{"type":9,"tag":32,"props":285,"children":286},{},[287],{"type":15,"value":288},"Important idea:",{"type":9,"tag":63,"props":290,"children":291},{},[292,297],{"type":9,"tag":67,"props":293,"children":294},{},[295],{"type":15,"value":296},"In large proteins, different relaxation pathways partially cancel.",{"type":9,"tag":67,"props":298,"children":299},{},[300],{"type":15,"value":301},"TROSY isolates the best one.",{"type":9,"tag":32,"props":303,"children":304},{},[305],{"type":15,"value":306},"Result:",{"type":9,"tag":63,"props":308,"children":309},{},[310,315],{"type":9,"tag":67,"props":311,"children":312},{},[313],{"type":15,"value":314},"Much sharper peaks",{"type":9,"tag":67,"props":316,"children":317},{},[318],{"type":15,"value":319},"Works well > 30 kDa",{"type":9,"tag":43,"props":321,"children":322},{},[],{"type":9,"tag":25,"props":324,"children":326},{"id":325},"page-9-trosy-deuteration",[327],{"type":15,"value":328},"Page 9 – TROSY + Deuteration",{"type":9,"tag":32,"props":330,"children":331},{},[332],{"type":15,"value":333},"Together → powerful.\nEven 45 kDa protein becomes feasible.",{"type":9,"tag":43,"props":335,"children":336},{},[],{"type":9,"tag":10,"props":338,"children":340},{"id":339},"_3-disordered-proteins-page-10",[341],{"type":15,"value":342},"🧬 3. Disordered Proteins (Page 10)",{"type":9,"tag":32,"props":344,"children":345},{},[346],{"type":15,"value":347},"α-synuclein vs calmodulin.",{"type":9,"tag":32,"props":349,"children":350},{},[351],{"type":15,"value":352},"Disordered proteins:",{"type":9,"tag":63,"props":354,"children":355},{},[356,361],{"type":9,"tag":67,"props":357,"children":358},{},[359],{"type":15,"value":360},"Narrow chemical shift dispersion",{"type":9,"tag":67,"props":362,"children":363},{},[364],{"type":15,"value":365},"All HN clustered around ~8 ppm",{"type":9,"tag":32,"props":367,"children":368},{},[369],{"type":15,"value":370},"Very heavy overlap.",{"type":9,"tag":43,"props":372,"children":373},{},[],{"type":9,"tag":10,"props":375,"children":377},{"id":376},"_4-sample-requirements-page-12",[378],{"type":15,"value":379},"🧪 4. Sample Requirements (Page 12)",{"type":9,"tag":32,"props":381,"children":382},{},[383],{"type":15,"value":384},"Critical:",{"type":9,"tag":63,"props":386,"children":387},{},[388,400,411,416,457],{"type":9,"tag":67,"props":389,"children":390},{},[391],{"type":9,"tag":392,"props":393,"children":394},"blockquote",{},[395],{"type":9,"tag":32,"props":396,"children":397},{},[398],{"type":15,"value":399},"1 mmol material",{"type":9,"tag":67,"props":401,"children":402},{},[403],{"type":9,"tag":392,"props":404,"children":405},{},[406],{"type":9,"tag":32,"props":407,"children":408},{},[409],{"type":15,"value":410},"0.2 mM concentration",{"type":9,"tag":67,"props":412,"children":413},{},[414],{"type":15,"value":415},"Stable for weeks",{"type":9,"tag":67,"props":417,"children":418},{},[419,421],{"type":15,"value":420},"Isotope labeling depends on size:",{"type":9,"tag":63,"props":422,"children":423},{},[424,435,446],{"type":9,"tag":67,"props":425,"children":426},{},[427],{"type":9,"tag":392,"props":428,"children":429},{},[430],{"type":9,"tag":32,"props":431,"children":432},{},[433],{"type":15,"value":434},"50 aa → ¹⁵N",{"type":9,"tag":67,"props":436,"children":437},{},[438],{"type":9,"tag":392,"props":439,"children":440},{},[441],{"type":9,"tag":32,"props":442,"children":443},{},[444],{"type":15,"value":445},"100 aa → ¹⁵N/¹³C",{"type":9,"tag":67,"props":447,"children":448},{},[449],{"type":9,"tag":392,"props":450,"children":451},{},[452],{"type":9,"tag":32,"props":453,"children":454},{},[455],{"type":15,"value":456},"300 aa → ²H/¹⁵N/¹³C",{"type":9,"tag":67,"props":458,"children":459},{},[460],{"type":15,"value":461},"Protein must be folded & monodisperse",{"type":9,"tag":32,"props":463,"children":464},{},[465],{"type":15,"value":466},"Without good sample → no NMR.",{"type":9,"tag":43,"props":468,"children":469},{},[],{"type":9,"tag":10,"props":471,"children":473},{"id":472},"_5-assignment-strategy-overview-page-13",[474],{"type":15,"value":475},"🧠 5. Assignment Strategy Overview (Page 13)",{"type":9,"tag":32,"props":477,"children":478},{},[479],{"type":15,"value":480},"Workflow:",{"type":9,"tag":482,"props":483,"children":484},"ol",{},[485,490,495,500,505],{"type":9,"tag":67,"props":486,"children":487},{},[488],{"type":15,"value":489},"Sample preparation",{"type":9,"tag":67,"props":491,"children":492},{},[493],{"type":15,"value":494},"Resonance assignment (huge time)",{"type":9,"tag":67,"props":496,"children":497},{},[498],{"type":15,"value":499},"NOE collection",{"type":9,"tag":67,"props":501,"children":502},{},[503],{"type":15,"value":504},"Structure calculation",{"type":9,"tag":67,"props":506,"children":507},{},[508],{"type":15,"value":509},"Functional/dynamics studies",{"type":9,"tag":32,"props":511,"children":512},{},[513],{"type":15,"value":514},"Assignment is foundation.",{"type":9,"tag":43,"props":516,"children":517},{},[],{"type":9,"tag":10,"props":519,"children":521},{"id":520},"_6-homonuclear-assignment-small-peptides-pages-1530",[522],{"type":15,"value":523},"🧭 6. Homonuclear Assignment – Small Peptides (Pages 15–30)",{"type":9,"tag":32,"props":525,"children":526},{},[527],{"type":15,"value":528},"Used when no isotope labeling.",{"type":9,"tag":18,"props":530,"children":532},{"id":531},"required-spectra-page-16",[533],{"type":15,"value":534},"Required spectra (Page 16):",{"type":9,"tag":63,"props":536,"children":537},{},[538,543,548],{"type":9,"tag":67,"props":539,"children":540},{},[541],{"type":15,"value":542},"TOCSY (80–90 ms)",{"type":9,"tag":67,"props":544,"children":545},{},[546],{"type":15,"value":547},"NOESY (50–60 ms)",{"type":9,"tag":67,"props":549,"children":550},{},[551],{"type":15,"value":552},"2QF-COSY",{"type":9,"tag":43,"props":554,"children":555},{},[],{"type":9,"tag":10,"props":557,"children":559},{"id":558},"_6a-understanding-cosy-vs-tocsy-pages-1719",[560],{"type":15,"value":561},"🧬 6A. Understanding COSY vs TOCSY (Pages 17–19)",{"type":9,"tag":25,"props":563,"children":565},{"id":564},"cosy",[566],{"type":15,"value":567},"🔹 COSY",{"type":9,"tag":32,"props":569,"children":570},{},[571,573,578],{"type":15,"value":572},"Shows ",{"type":9,"tag":110,"props":574,"children":575},{},[576],{"type":15,"value":577},"direct J-couplings only",{"type":15,"value":579}," (usually 2–3 bonds).",{"type":9,"tag":32,"props":581,"children":582},{},[583],{"type":15,"value":584},"So:\nHα ↔ Hβ\nHβ ↔ Hγ",{"type":9,"tag":32,"props":586,"children":587},{},[588],{"type":15,"value":589},"But NOT:\nHα ↔ Hγ (unless directly coupled)",{"type":9,"tag":32,"props":591,"children":592},{},[593],{"type":15,"value":594},"COSY = nearest neighbors in bond network.",{"type":9,"tag":43,"props":596,"children":597},{},[],{"type":9,"tag":25,"props":599,"children":601},{"id":600},"tocsy",[602],{"type":15,"value":603},"🔹 TOCSY",{"type":9,"tag":32,"props":605,"children":606},{},[607,608,613],{"type":15,"value":572},{"type":9,"tag":110,"props":609,"children":610},{},[611],{"type":15,"value":612},"entire spin system",{"type":15,"value":614}," via relayed magnetization.",{"type":9,"tag":32,"props":616,"children":617},{},[618],{"type":15,"value":619},"All protons within same residue connect.",{"type":9,"tag":32,"props":621,"children":622},{},[623],{"type":15,"value":624},"Example:\nHN → Hα → Hβ → Hγ → Hδ → etc.",{"type":9,"tag":32,"props":626,"children":627},{},[628],{"type":15,"value":629},"TOCSY = entire amino acid fingerprint.",{"type":9,"tag":43,"props":631,"children":632},{},[],{"type":9,"tag":10,"props":634,"children":636},{"id":635},"_6b-what-is-a-spin-system-page-17",[637],{"type":15,"value":638},"🧬 6B. What is a Spin System? (Page 17)",{"type":9,"tag":32,"props":640,"children":641},{},[642,644],{"type":15,"value":643},"Important:\n",{"type":9,"tag":110,"props":645,"children":646},{},[647],{"type":15,"value":648},"No J-coupling across peptide bond.",{"type":9,"tag":32,"props":650,"children":651},{},[652],{"type":15,"value":653},"Each amino acid = independent spin system.",{"type":9,"tag":32,"props":655,"children":656},{},[657],{"type":15,"value":658},"This is the key assumption for assignment.",{"type":9,"tag":43,"props":660,"children":661},{},[],{"type":9,"tag":10,"props":663,"children":665},{"id":664},"_6c-sequential-assignment-procedure-pages-2028",[666],{"type":15,"value":667},"🧬 6C. Sequential Assignment Procedure (Pages 20–28)",{"type":9,"tag":25,"props":669,"children":671},{"id":670},"step-1-number-hnhα-peaks-page-2022",[672],{"type":15,"value":673},"Step 1 – Number HN/Hα peaks (Page 20–22)",{"type":9,"tag":32,"props":675,"children":676},{},[677],{"type":15,"value":678},"Arbitrarily label them.",{"type":9,"tag":25,"props":680,"children":682},{"id":681},"step-2-use-tocsy",[683],{"type":15,"value":684},"Step 2 – Use TOCSY",{"type":9,"tag":32,"props":686,"children":687},{},[688],{"type":15,"value":689},"Find all peaks belonging to same spin system.",{"type":9,"tag":25,"props":691,"children":693},{"id":692},"step-3-identify-residue-type-page-23",[694],{"type":15,"value":695},"Step 3 – Identify residue type (Page 23)",{"type":9,"tag":32,"props":697,"children":698},{},[699],{"type":15,"value":700},"Examples:",{"type":9,"tag":63,"props":702,"children":703},{},[704,709,714,719,724,729,734],{"type":9,"tag":67,"props":705,"children":706},{},[707],{"type":15,"value":708},"Gly: 2 Hα, nothing else",{"type":9,"tag":67,"props":710,"children":711},{},[712],{"type":15,"value":713},"Ala: Hα + CH₃",{"type":9,"tag":67,"props":715,"children":716},{},[717],{"type":15,"value":718},"Thr: 2 Hα-like + CH₃",{"type":9,"tag":67,"props":720,"children":721},{},[722],{"type":15,"value":723},"Val: Hα + Hβ + 2 CH₃",{"type":9,"tag":67,"props":725,"children":726},{},[727],{"type":15,"value":728},"AMX pattern: Ser, Cys, Asp, Asn, His, Phe, Tyr, Trp",{"type":9,"tag":67,"props":730,"children":731},{},[732],{"type":15,"value":733},"Lys/Arg: many signals 3–3.5 ppm",{"type":9,"tag":67,"props":735,"children":736},{},[737],{"type":15,"value":738},"Pro: no HN → breaks walk",{"type":9,"tag":32,"props":740,"children":741},{},[742],{"type":15,"value":743},"This is pattern recognition.",{"type":9,"tag":43,"props":745,"children":746},{},[],{"type":9,"tag":10,"props":748,"children":750},{"id":749},"_6d-sequential-noe-walk-page-26",[751],{"type":15,"value":752},"🧬 6D. Sequential NOE Walk (Page 26)",{"type":9,"tag":32,"props":754,"children":755},{},[756],{"type":15,"value":757},"Critical rule:",{"type":9,"tag":32,"props":759,"children":760},{},[761],{"type":15,"value":762},"HN(i) shows NOE to:",{"type":9,"tag":63,"props":764,"children":765},{},[766,771],{"type":9,"tag":67,"props":767,"children":768},{},[769],{"type":15,"value":770},"Hα(i-1)",{"type":9,"tag":67,"props":772,"children":773},{},[774],{"type":15,"value":775},"Hβ(i-1)",{"type":9,"tag":32,"props":777,"children":778},{},[779],{"type":15,"value":780},"NEVER forward.",{"type":9,"tag":32,"props":782,"children":783},{},[784],{"type":15,"value":785},"So you connect:\nResidue i ↔ residue i-1",{"type":9,"tag":32,"props":787,"children":788},{},[789],{"type":15,"value":790},"This is the “sequential walk”.",{"type":9,"tag":43,"props":792,"children":793},{},[],{"type":9,"tag":25,"props":795,"children":797},{"id":796},"important-note",[798],{"type":15,"value":799},"Important note:",{"type":9,"tag":32,"props":801,"children":802},{},[803],{"type":15,"value":804},"HN-HN NOEs appear only in α-helices.",{"type":9,"tag":32,"props":806,"children":807},{},[808],{"type":15,"value":809},"In β-sheets → interstrand NOEs appear (Page 31).",{"type":9,"tag":32,"props":811,"children":812},{},[813],{"type":15,"value":814},"These can confuse assignment.",{"type":9,"tag":43,"props":816,"children":817},{},[],{"type":9,"tag":10,"props":819,"children":821},{"id":820},"️-pitfalls-page-30",[822],{"type":15,"value":823},"⚠️ Pitfalls (Page 30)",{"type":9,"tag":63,"props":825,"children":826},{},[827,832,837,842],{"type":9,"tag":67,"props":828,"children":829},{},[830],{"type":15,"value":831},"Overlap",{"type":9,"tag":67,"props":833,"children":834},{},[835],{"type":15,"value":836},"Prolines break walk",{"type":9,"tag":67,"props":838,"children":839},{},[840],{"type":15,"value":841},"Secondary structure gives extra NOEs",{"type":9,"tag":67,"props":843,"children":844},{},[845],{"type":15,"value":846},"Very difficult without labeling",{"type":9,"tag":43,"props":848,"children":849},{},[],{"type":9,"tag":10,"props":851,"children":853},{"id":852},"_7-n-labeling-only-pages-3233",[854],{"type":15,"value":855},"🧪 7. ¹⁵N Labeling Only (Pages 32–33)",{"type":9,"tag":32,"props":857,"children":858},{},[859],{"type":15,"value":860},"Use ¹H–¹⁵N HSQC.",{"type":9,"tag":32,"props":862,"children":863},{},[864],{"type":15,"value":865},"Each amide = one peak.",{"type":9,"tag":32,"props":867,"children":868},{},[869],{"type":15,"value":870},"Better dispersion because ¹⁵N axis spreads signals.",{"type":9,"tag":43,"props":872,"children":873},{},[],{"type":9,"tag":18,"props":875,"children":877},{"id":876},"_3d-n-edited-tocsy-page-33",[878],{"type":15,"value":879},"🔹 3D ¹⁵N-edited TOCSY (Page 33)",{"type":9,"tag":32,"props":881,"children":882},{},[883],{"type":15,"value":884},"Dimensions:",{"type":9,"tag":482,"props":886,"children":887},{},[888,893,898],{"type":9,"tag":67,"props":889,"children":890},{},[891],{"type":15,"value":892},"HN",{"type":9,"tag":67,"props":894,"children":895},{},[896],{"type":15,"value":897},"¹⁵N",{"type":9,"tag":67,"props":899,"children":900},{},[901],{"type":15,"value":902},"side-chain proton",{"type":9,"tag":32,"props":904,"children":905},{},[906],{"type":15,"value":907},"Interpretation:\nTake one HSQC peak → look into 3rd dimension → see full spin system.",{"type":9,"tag":32,"props":909,"children":910},{},[911],{"type":15,"value":912},"This removes overlap in HN dimension.",{"type":9,"tag":32,"props":914,"children":915},{},[916],{"type":15,"value":917},"Limitation:\nWeak for larger proteins.",{"type":9,"tag":43,"props":919,"children":920},{},[],{"type":9,"tag":10,"props":922,"children":924},{"id":923},"_8-triple-resonance-experiments-pages-3441",[925],{"type":15,"value":926},"🧪 8. Triple Resonance Experiments (Pages 34–41)",{"type":9,"tag":32,"props":928,"children":929},{},[930],{"type":15,"value":931},"This is the modern method.",{"type":9,"tag":32,"props":933,"children":934},{},[935],{"type":15,"value":936},"Requires ¹⁵N/¹³C labeling.",{"type":9,"tag":43,"props":938,"children":939},{},[],{"type":9,"tag":18,"props":941,"children":943},{"id":942},"hnca-vs-hncoca-pages-3539",[944],{"type":15,"value":945},"🔹 HNCA vs HN(CO)CA (Pages 35–39)",{"type":9,"tag":25,"props":947,"children":949},{"id":948},"hnca-detects",[950],{"type":15,"value":951},"HNCA detects:",{"type":9,"tag":32,"props":953,"children":954},{},[955],{"type":15,"value":956},"Cα(i) AND Cα(i-1)",{"type":9,"tag":25,"props":958,"children":960},{"id":959},"hncoca-detects",[961],{"type":15,"value":962},"HN(CO)CA detects:",{"type":9,"tag":32,"props":964,"children":965},{},[966],{"type":15,"value":967},"ONLY Cα(i-1)",{"type":9,"tag":32,"props":969,"children":970},{},[971],{"type":15,"value":972},"So:",{"type":9,"tag":32,"props":974,"children":975},{},[976],{"type":15,"value":977},"If peak appears in both → Cα(i-1)\nIf only in HNCA → Cα(i)",{"type":9,"tag":32,"props":979,"children":980},{},[981],{"type":15,"value":982},"This is the backbone linking logic.",{"type":9,"tag":43,"props":984,"children":985},{},[],{"type":9,"tag":18,"props":987,"children":989},{"id":988},"hnco-hncaco",[990],{"type":15,"value":991},"🔹 HNCO / HN(CA)CO",{"type":9,"tag":32,"props":993,"children":994},{},[995],{"type":15,"value":996},"Same idea but for carbonyl (C').",{"type":9,"tag":43,"props":998,"children":999},{},[],{"type":9,"tag":18,"props":1001,"children":1003},{"id":1002},"cbcaconh-cbcanh",[1004],{"type":15,"value":1005},"🔹 CBCA(CO)NH / CBCANH",{"type":9,"tag":32,"props":1007,"children":1008},{},[1009],{"type":15,"value":1010},"Same logic for Cα and Cβ.",{"type":9,"tag":32,"props":1012,"children":1013},{},[1014],{"type":15,"value":1015},"Gives amino acid identity.",{"type":9,"tag":43,"props":1017,"children":1018},{},[],{"type":9,"tag":10,"props":1020,"children":1022},{"id":1021},"_9-how-to-interpret-3d-spectra-page-37-strips",[1023],{"type":15,"value":1024},"🧲 9. How to Interpret 3D Spectra (Page 37 – Strips)",{"type":9,"tag":32,"props":1026,"children":1027},{},[1028],{"type":15,"value":1029},"A 3D spectrum has 3 axes:",{"type":9,"tag":32,"props":1031,"children":1032},{},[1033],{"type":15,"value":1034},"Example:",{"type":9,"tag":63,"props":1036,"children":1037},{},[1038,1043,1047],{"type":9,"tag":67,"props":1039,"children":1040},{},[1041],{"type":15,"value":1042},"¹H",{"type":9,"tag":67,"props":1044,"children":1045},{},[1046],{"type":15,"value":897},{"type":9,"tag":67,"props":1048,"children":1049},{},[1050],{"type":15,"value":1051},"¹³C",{"type":9,"tag":32,"props":1053,"children":1054},{},[1055],{"type":15,"value":1056},"Visualization method:\nYou slice at one HSQC peak → generate a “strip”.",{"type":9,"tag":32,"props":1058,"children":1059},{},[1060],{"type":15,"value":1061},"Each strip:\n= one residue",{"type":9,"tag":32,"props":1063,"children":1064},{},[1065],{"type":15,"value":1066},"Then compare neighboring strips:\nFind matching Cα(i-1) to Cα(i)",{"type":9,"tag":32,"props":1068,"children":1069},{},[1070],{"type":15,"value":1071},"That builds sequence.",{"type":9,"tag":32,"props":1073,"children":1074},{},[1075],{"type":15,"value":1076},"Strip comparison is like matching puzzle pieces.",{"type":9,"tag":43,"props":1078,"children":1079},{},[],{"type":9,"tag":10,"props":1081,"children":1083},{"id":1082},"_10-chemical-shift-patterns-page-42",[1084],{"type":15,"value":1085},"🧪 10. Chemical Shift Patterns (Page 42)",{"type":9,"tag":32,"props":1087,"children":1088},{},[1089],{"type":15,"value":1090},"Cα vs Cβ shifts cluster by residue type.",{"type":9,"tag":32,"props":1092,"children":1093},{},[1094],{"type":15,"value":1034},{"type":9,"tag":63,"props":1096,"children":1097},{},[1098,1103,1108],{"type":9,"tag":67,"props":1099,"children":1100},{},[1101],{"type":15,"value":1102},"Gly: low Cα",{"type":9,"tag":67,"props":1104,"children":1105},{},[1106],{"type":15,"value":1107},"Thr: high Cβ",{"type":9,"tag":67,"props":1109,"children":1110},{},[1111],{"type":15,"value":1112},"Val/Ile: characteristic pattern",{"type":9,"tag":32,"props":1114,"children":1115},{},[1116],{"type":15,"value":1117},"This helps residue identification.",{"type":9,"tag":43,"props":1119,"children":1120},{},[],{"type":9,"tag":10,"props":1122,"children":1124},{"id":1123},"_11-side-chain-assignment-pages-4345",[1125],{"type":15,"value":1126},"🧬 11. Side Chain Assignment (Pages 43–45)",{"type":9,"tag":32,"props":1128,"children":1129},{},[1130],{"type":15,"value":1131},"Experiments:",{"type":9,"tag":63,"props":1133,"children":1134},{},[1135,1140,1145,1150],{"type":9,"tag":67,"props":1136,"children":1137},{},[1138],{"type":15,"value":1139},"H(C)CH-TOCSY",{"type":9,"tag":67,"props":1141,"children":1142},{},[1143],{"type":15,"value":1144},"(H)CCH-TOCSY",{"type":9,"tag":67,"props":1146,"children":1147},{},[1148],{"type":15,"value":1149},"H(CCCO)NH",{"type":9,"tag":67,"props":1151,"children":1152},{},[1153],{"type":15,"value":1154},"HCCH-type",{"type":9,"tag":32,"props":1156,"children":1157},{},[1158],{"type":15,"value":1159},"These transfer magnetization across carbon network.",{"type":9,"tag":32,"props":1161,"children":1162},{},[1163],{"type":15,"value":1164},"Used for:",{"type":9,"tag":63,"props":1166,"children":1167},{},[1168,1173],{"type":9,"tag":67,"props":1169,"children":1170},{},[1171],{"type":15,"value":1172},"Aliphatic side chains",{"type":9,"tag":67,"props":1174,"children":1175},{},[1176],{"type":15,"value":1177},"Aromatics (very difficult)",{"type":9,"tag":32,"props":1179,"children":1180},{},[1181],{"type":15,"value":1182},"Assignment often takes 50% of project time.",{"type":9,"tag":43,"props":1184,"children":1185},{},[],{"type":9,"tag":10,"props":1187,"children":1189},{"id":1188},"_12-complete-experiment-set-page-46",[1190],{"type":15,"value":1191},"🧾 12. Complete Experiment Set (Page 46)",{"type":9,"tag":32,"props":1193,"children":1194},{},[1195],{"type":15,"value":1196},"Typical backbone set:",{"type":9,"tag":63,"props":1198,"children":1199},{},[1200,1205,1210,1215,1220,1225,1230,1235],{"type":9,"tag":67,"props":1201,"children":1202},{},[1203],{"type":15,"value":1204},"HNCA",{"type":9,"tag":67,"props":1206,"children":1207},{},[1208],{"type":15,"value":1209},"HN(CO)CA",{"type":9,"tag":67,"props":1211,"children":1212},{},[1213],{"type":15,"value":1214},"HNCO",{"type":9,"tag":67,"props":1216,"children":1217},{},[1218],{"type":15,"value":1219},"HN(CA)CO",{"type":9,"tag":67,"props":1221,"children":1222},{},[1223],{"type":15,"value":1224},"CBCA(CO)NH",{"type":9,"tag":67,"props":1226,"children":1227},{},[1228],{"type":15,"value":1229},"CBCANH",{"type":9,"tag":67,"props":1231,"children":1232},{},[1233],{"type":15,"value":1234},"HBHA(CBCACO)NH",{"type":9,"tag":67,"props":1236,"children":1237},{},[1238],{"type":15,"value":1239},"HCCH-TOCSY",{"type":9,"tag":32,"props":1241,"children":1242},{},[1243],{"type":15,"value":1244},"Exact choice depends on protein size.",{"type":9,"tag":43,"props":1246,"children":1247},{},[],{"type":9,"tag":10,"props":1249,"children":1251},{"id":1250},"_13-literature-assignments-page-47",[1252],{"type":15,"value":1253},"📚 13. Literature Assignments (Page 47)",{"type":9,"tag":32,"props":1255,"children":1256},{},[1257,1259],{"type":15,"value":1258},"BioMagResBank (BMRB)\n",{"type":9,"tag":1260,"props":1261,"children":1265},"a",{"href":1262,"rel":1263},"http://www.bmrb.wisc.edu",[1264],"nofollow",[1266],{"type":15,"value":1262},{"type":9,"tag":32,"props":1268,"children":1269},{},[1270],{"type":15,"value":1271},"Contains deposited assignments.",{"type":9,"tag":43,"props":1273,"children":1274},{},[],{"type":9,"tag":10,"props":1276,"children":1278},{"id":1277},"big-picture-summary",[1279],{"type":15,"value":1280},"🎯 Big Picture Summary",{"type":9,"tag":32,"props":1282,"children":1283},{},[1284],{"type":15,"value":1285},"Protein NMR challenges:",{"type":9,"tag":482,"props":1287,"children":1288},{},[1289,1294,1299],{"type":9,"tag":67,"props":1290,"children":1291},{},[1292],{"type":15,"value":1293},"Signal overlap",{"type":9,"tag":67,"props":1295,"children":1296},{},[1297],{"type":15,"value":1298},"T₂ relaxation",{"type":9,"tag":67,"props":1300,"children":1301},{},[1302],{"type":15,"value":1303},"Large number of resonances",{"type":9,"tag":32,"props":1305,"children":1306},{},[1307],{"type":15,"value":1308},"Solutions:",{"type":9,"tag":63,"props":1310,"children":1311},{},[1312,1317,1322,1327,1332],{"type":9,"tag":67,"props":1313,"children":1314},{},[1315],{"type":15,"value":1316},"2D/3D experiments",{"type":9,"tag":67,"props":1318,"children":1319},{},[1320],{"type":15,"value":1321},"Isotope labeling",{"type":9,"tag":67,"props":1323,"children":1324},{},[1325],{"type":15,"value":1326},"Deuteration",{"type":9,"tag":67,"props":1328,"children":1329},{},[1330],{"type":15,"value":1331},"TROSY",{"type":9,"tag":67,"props":1333,"children":1334},{},[1335],{"type":15,"value":1336},"Triple resonance backbone linking",{"type":9,"tag":32,"props":1338,"children":1339},{},[1340],{"type":15,"value":1341},"Assignment logic:",{"type":9,"tag":32,"props":1343,"children":1344},{},[1345],{"type":15,"value":1346},"Spin system → sequential connection → backbone linking → side chain completion → NOEs → structure.",1775084299381]