Protein-Protein interactionics

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Protein-Protein interactionics

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• ????????????????????????????????????????????????

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• 1.?????????????????????
• 2.?????????????????????
• 3.????????????????
• 4.????????????????
• 5.???????????????????
• 6.?????????????????
• 7.????????????????

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• ??????(Labeled fusion protein)
• ?????(co-immunoprecipitation)
• ?????(Yeast two hybrid system)
• ??????????(FRET)
• ?????????(Biacore SPR??)
• ????????(Atomic force microscopy)
• ???????(Phage display)

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• Received 11 January 2008
• Received in revised form 14 March 2008
• Accepted 21 April 2008
• Available online 29 April 2008

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• The detection of proteinprotein binding on
  microarrays using the fluorescence lifetime as a
  dynamic analytical parameter was investigated in
  a model system. The assay is based on Forster
  resonance energy transfer (FRET) and carried out
  with biotinylated Bovine Seru Albumin and
  streptavidin, labeled with the commonly used
  microarray dyes Alexa 555 and Alexa 647,
  respectively. This efficient FRET donor/acceptor
  pairwas employed in a competitive assay format on
  three different microarray surfaceson CEL Epoxy,
  ARChip Epoxy and ARChip Gel surfaces .Lifetime
  maps were recorded according to the Rapid
  Lifetime Determination (RLD) scheme. Good
  linearity of the quenching curve was obtained in
  all cases.

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FRET????

• FRET??????????????????????????????????????????????
  ???,???????????????????,???????????????,??????????
  ??????????????????????
• ??????????,FRET????????????????,?????????,????????
  ?????????FRET?FRIM????????????????????????????????
  ?????????????????

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??1
??2
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Fig. 2. Lifetime maps of Alexa 555-BSA-Biotin
spots incubated with various amounts of
streptavidin on three different surfaces
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Fig. 3. Average lifetimes of Alexa 555-BSA-Biotin
after incubation with streptavidin on (a) CEL
Epoxy slides, (b) ARChip Epoxy slides, and (c)
ARChip Gel slides and linear fits.
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• ???surfaces?????streptavidin?,????????????,???????
  ??????
• ?streptavidin???100nM?,?? surfaces????????????????
  ,?Epoxy surfaces???,??????????????
• ??surfaces, streptavidin???????????????????,?Cel
  surfaces????,???????????
• ???????,?????????????????,?????????

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FRET??????

• ??
• 1.good precision, sensitivity and little
• susceptibility for interferences,a high degree
  of flexibility and multiexponential
• 2.??????????????????-??????????????
• ????????????

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?????????FRET????

• W-FRET ???????FRET???
• C-FRET??????-FRET???
• Confocal- FRET???????-FRET???
• MP-FRET ???-FRET???

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• ?????????????(BIND)
• ?????????(DIP)
• ??????????JSP??http//point.bioinformatics.tw/intr
  o/intro.jsp

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• Thank you!
• The questions are appreciated!