具有高通量CD的多变量二级结构估计（SSE）

智能文字提取功能测试中

CD的测量揭示了关于蛋白质的二级结构或主干构象的信息。生物分子的二级结构含量的测定可以阐明结构和功能之间的关系或验证蛋白质的稳定性。JASCO的多元二级结构估计（SSE）程序可用于分析CD数据，以便为研究人员提供更多的定量结构估计。使用高通量CD（HTCD）系统和多元SSE程序，可以自动测量和有效地分析大量的样本。本应用说明说明了在CD测量后获得的二级结构数据中的使用。PLS方法与x射线晶体学得到的二级结构结果吻合较好。Application NoteCDSSpectroscopy Using the Multivariate Secondary StructureEstimation (SSE) program2/4Application Note Using the Multivariate Secondary Structure Estimation (SSE) program Introduction CD m ea s uremen t s revea l i n formation regard in g the s e c on d a r y str u ctu r e o r ba c k b one co n f o rm a t i o n of a pro t ei n. D e t e rmin ati o n of the s ec o n da r y stru c tu r e co n ten t of a b i omole c u l e ca n e lu cidate the relationship between st ru c tur e an d func t ion o r v e r ify protei n st a b i lity . J A SCO's mu ltiv a ri a t e S eco n dar y St ru c tu r e E s t i m a t ion (SS E) pro g r am ca n be used t o a n a l yse CD data in o r de r to s u pp l y r es e a rc h e rs w i t h more q u a n ti t a tiv e s t ru c ture e s t i mation s . A l a rge nu mb e r of sa mpl es c a n be au t o mati ca lly m e a s ur ed an d eff i cie n tly analysed u si ng the H i g h -Th r o u ghpu t CD (HTC D ) s y s tem and mu lti var i a t e SSE pro g ra m. JASCO J-1500 high-throughput CD system View product information at w ww.j a sc oin c.c o m Th i s a ppl i c a ti o n no te s dem o n st ra t es s eco n da r y st ru c tu r e res u lts o f e ig h t pro t ein s u s i n g th e M u lti va ri a te SSE pr o g ram. Keywords Second a ry stru c tu r e estimation， circ u la r d ichro i sm， H TCD，M u ltiv a ri a te ， B i oc h e m is tr y， Pha rma ce u ticals Experimental Measurement conditions D a t a acq ui siti o n in t e rv a l 0.1n m R esp o nse t im e 2 sec o n d s S p ect ra l ba n dw idt h 1n m Sc an s p ee d 100 nm /min Acc u m u l a tions 2 ti mes Path lengt h 1mm Results Prior t o obta inin g the s eq u e n ce me a su r eme n t ， the path le n gth and m e a n r esidue mol ar conce n trat i o n s h a ve to be speci f ied s o th at t he o pti c al c on st an t s c an b e au t o mati ca lly c a l c u la t e d f or the s ec o n da r y s tru ct ure est im a ti o n (Figur e 1) Figure 1. Multivariate parameter setup speci f ying path length and mean residue molar concentration The CD me a s u r eme n t s in the fa r-UV reg i on a r e o bt a in ed fo r e i g h t 0.1 mg /mL protein s a mp l es wit h va r y i n g se c onda r y st ru c tu re co m pos i ti o n s an d are sh o w n in F ig ur e 2. Figure 2. CD spectra of protein samples Th e secondary stru ct ure is estimated from the prote in spec tr a in Figure 1 u s ing the PL S mul t i v ariate met h od. Table co mp a r es these r esu lt s wi th X-ray c r ystallo g raphy d ata. F i gu r e 3 shows th e seco nd a r y str u ct u r e r esu lt s set u p i n the JWM V S p r o gra m. 28600 Mary's Cour t ， E asto n ， MD 21601US A Table 1. Comparison of Mult i variate SSE results with X-ray crystallography data a-Helix % B-Sheet% Turn% Other % Lysozyme PLS 42.8 0.4 24.4 32.4 X-ray' 41 4 19 35 Cytochrome C PLS 42.6 3.1 18.1 36.2 X-ray' 42 8 9 42 Concanavalin A PLS 5.1 44.6 13.9 36.4 X-ray' 2 36 12 49 B-Lactoglobulin PLS 17.8 35.5 12.3 34.4 X-ray' 13 34 13 41 Trypsin inhibitor PLS 13.9 25.3 17.3 43.5 X-ray 2 33 10 55 Ribonuclease A PLS 21.5 14.7 22.4 41.4 X-ray' 22 19 11 48 Human SerumAlbumin (HSA) PLS 66.8 1.3 8.2 23.7 X-ray² 72 0 8 29 Hemoglobin PLS 61.1 0 18 20.9 X-ray' 75 0 10 15 W. C. Johnson， Proteins： Structure， Function， and Genetics (1999)， 35，307-312. Figure 3. Results set up using the Mult i variate SSE program Conclusion This application n ote i llu s tr a t es the u se o f the Mu lti va ri a t e SSE program i n obtained seco n dary s t r uc tu re d ata aftei C D m e a su r em en t s . T h e PL S metho d sho w s g o od agr ee m en t wi th seco n dar y st r uc tu re res u lts o bt a in e d by X -ra y c ry s tallog r a ph y. 28600 Mary's Cour t ， E asto n ， MD 21601US A