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Development of peptide chip for rough diagnosis

Development of peptide chip for rough diagnosis. Deguchi Nao E-mail address : ac1210nd@apps.kct.ac.jp Department of Material Sciences & Chemical Engineering, Kitakyushu National College of Technology. introduction.

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Development of peptide chip for rough diagnosis

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  1. Development of peptide chip for rough diagnosis DeguchiNao E-mail address : ac1210nd@apps.kct.ac.jp Department of Material Sciences & Chemical Engineering, Kitakyushu National College of Technology

  2. introduction ・Severe illness that are difficult to cure cause significant impact on our daily lives. ・However, with early detection, it is quite possible to slow the progression, or even cure the disease with appropriate treatment. As a method to easily diagnose these diseases, I have been developing a peptide chip that could potentially be a break-thorough for simple diagnostic methodologies.

  3. It is understood that the majority of the causes of untreatable diseases originate in some kinds of abnormality in the intracellular signal transduction system. If there are methodologies to which we can monitor these intercellular signal transductions, diagnosis of these untreatable diseases will be possible. however it is practically impossible to completely understand the intracellular signal transductions. Protein phosphorylation signaling which is one of the most important and versatile intracellular signaling methods

  4. protein phosphorylation －a reaction of the g-phosphate group of ATP transferring to a hydroxy group at the side chain of serine, threonine and tyrosine residues, which are included in substrate peptides or proteins in cells. 200 kinds of protein kinase, which is an enzyme catalyzing phosphorylation reaction, in human body. In order to monitor all protein kinase activities at once, I have tried to develop a analytical technique utilizing peptide array and MALDI-TOF-MS.

  5. principle of MALDI-TOF-MS + - photocleavagecompound detector P P P P substratepeptide substrate mass of phosphorylated peptide is increased by 80 rate of phosphorylation of MS spectra

  6. purpose In order to conduct mass spectrometry using a peptide array, Our team needs substrate peptides with the photo-cleavable part by ionization laser synthesis 2-bromo-2-(2-nitrophenyl)acetic acid (BNPA)

  7. method ●BNPA N-bromosuccinimide(6.0 g) + CCl4 (25 ml) + catalytic amountof benzoyl peroxide 2-(2-nitrophenyl)acetic acid (5.0 g) + thionylchloride (8ml) In carbon tetrachloride (5 ml) 65℃ 1.5ｈ 1.0ｈ 75℃ 4.5ｈ recrystallization from CH2Cl2 extract 3 x 25 ml CH2Cl2 ice(25 g)

  8. + + result ・shape: Solid needle ・color: Clear ・mp: 59.6 ~ 63.0 ℃ Table. mass spectrometry Figure.1H-NMR

  9. conclusion Immobilized concept substrate substrate I've tried the synthesis of the photo-cleavable compound.The synthesized one was identified with BNPA by 1H-NMR and mass spectrometry.

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