Protein Misfolding Amyloid Lab Report Example | Topics and Well Written Essays - 3000 words

Protein Misfolding Amyloid - science laboratory Report Example name 3. Fluorescence intensities of a) Tyrosine b) Tryptophan and c) Thioflavin T in Con A , at 40 0C a)b)c)Fig 3 b also shows considerable Trp fluorescence at pH 5.0 and 7.0 go at pH 9.0, mass remained insignificant again showing buried/ quenched Trp residues.After a lag of 40 h the ThT intensity change magnitude sharply to high level at pH 5.0. At pH 7.0, the plateau was attained later on slight increase up to 20 h. At pH 9.0 the intensity increased at 70 h.Amyloid A40 Assay Fig 4. Fluorescence of a) Tyrosine and b) Thioflavin T in A40 at 250 CFig 4 a)The ThioflavinT bound to A increases at 40 h decreases there afterward and increases indicating conformational changes.Fig 4 b)Fig 4. shows Tyr intensity decreasing after 60h and so is the Th T intensity however the last mentioned increases thereafter showing increased amyloidal filament formation but at this time the Tyr seems to be quenched somewhat.Fig 5. TE Mi crographs of Con A in pH 5.0 at 0 hFig 5 a) X10K some amorphous aggregates are already present at this pH Fig 6. TEM showing Con A in pH 5.0 at a) 4 (X40K) and, b) 48h (X40K) c) 48 h at pH 7.0 (X15K and d) 48h at pH 9.0 at 25 0C Fig 6 a) only amorphous aggregates are present Fig 6 b) Fibril formation is clearly visible.Fig 6 c) 48h at pH 7.0 (X15K)Fig 6 d) 48 h at pH 9.0 (X10K) as expected the long amyloid fibrils are in state of formationFig 7. Con A at 37.2 0C and pH 5.0 after 24h The physiological temperature and low pH shows abundant short rodsFig 8. A40 at 25 0 C a) 0, and b) 3h. (X20K) neither shows fibrils despite positive ThT fluorescence.Fig 7 shows AB40 as small...But at pH 9.0 the intensity diminish sharply from beginning up to 70h and then attained a plateau. The decrease of 400u indicated huge conformational change atomic number 82 to buried and /or quenched Trp residues.Th T fluorescence decreased slightly up to 50 hrs and increased sharply thereafter reaching at pe ak at 80 hrs and then decreasing considerably. At pH 7 the intensity increased from beginning, reaching a max at 50h and decreasing sharply, thereafter. The Th T fluorescence shows reversible trend in these experiments and conformational changes are occurring fast. At pH 9.0, there was considerable increase in fluorescence intensity after 75h showing fibrillation (fig 1 c).Highest intensity was observed at pH 5.0 while considerably high Intensity at pH 7.0. However at pH 9.0 there was minimum intensity for tyrosine. The latter temperature and pH combination either create conditions for Tyrosine quenching or the aggregates deeply disguise this amino acid as a result of conformational changes (Fig 3a).Fig 4. shows Tyr intensity decreasing after 60h and so is the Th T intensity however the latter increases thereafter showing increased amyloidal fibril formation but at this time the Tyr seems to be quenched somewhat.Only large amorphous structures seen after 72 h while a solitary long fibril is seen in the upper contribution after 96h (Fig 8 b).