Photo-reactive amino acid analogs are artificial analogs of natural amino acids that can be used for crosslinking of protein complexes. Photo-reactive amino acid analogs may be incorporated into proteins and peptides in vivo or in vitro. Photo-reactive amino acid analogs in common use are photoreactive diazirine analogs to leucine and methionine, and para-benzoylphenylalanine. Upon exposure to ultraviolet light, they are activated and covalently bind to interacting proteins that are within a few angstroms of the photo-reactive amino acid analog.

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dbo:abstract
  • Un aminoàcid anàleg fotoreactiu és un tipus d'aminoàcid artificial, anàleg als naturals, que pot ésser incorporat a proteïnes i pèptids amb la principal finalitat de detectar interaccions entre diverses proteïnes. Aquestes interaccions entre proteïnes són de vital importància per als processos cel·lulars (metabolisme, comunicació...) que es desenvolupen a l'interior dels organismes vius. Des de l'era genòmica va sorgir la important necessitat de saber quines són les organitzacions i xarxes proteiques que controlen les funcions cel·lulars, i són molts els estudis que han demostrat que la major part de les proteïnes funcionen com a complexos: interaccionant entre elles -reticulació- i no pas com a unitats individuals i separades. Uns exemples d'aquests aminoàcids fotoreactius són: la fotometionina -amb fórmula molecular C6H11N3O2- que deriva de la metionina (C5H11NO2S), o la fotoleucina (C5H9N3O2) que deriva de la leucina (C6H13NO2). Per tant, detectar aquestes interaccions i relacions entre les proteïnes serà molt important; no només per la investigació bàsica sinó també pel desenvolupament de tractaments i fàrmacs. (ca)
  • Photo-reactive amino acid analogs are artificial analogs of natural amino acids that can be used for crosslinking of protein complexes. Photo-reactive amino acid analogs may be incorporated into proteins and peptides in vivo or in vitro. Photo-reactive amino acid analogs in common use are photoreactive diazirine analogs to leucine and methionine, and para-benzoylphenylalanine. Upon exposure to ultraviolet light, they are activated and covalently bind to interacting proteins that are within a few angstroms of the photo-reactive amino acid analog. L-Photo-Leucine and L-Photo-Methionine are analogs of the naturally occurring L-Leucine and L-Methionine amino acids that are endogenously incorporated into the primary sequence of proteins during synthesis using the normal translation machinery. They are then ultraviolet light (UV)-activated to covalently crosslink proteins within protein–protein interaction domains in their native in-vivo environment. The method enables the determination and characterization of both stable and transient protein interactions in cells without the addition of chemical crosslinkers and associated solvents that can adversely affect the cell biology being studied in the experiment. When used in combination with limiting media that is devoid of leucine and methionine, the photo-activatable derivatives are treated like naturally occurring amino acids by the cellular protein synthesis machinery. As a result, they can be substituted for leucine or methionine in the primary structure of proteins. Photo-leucine and photo-methionine derivatives contain diazirine rings that are activated when exposed to UV light to become reactive intermediates that form covalent bonds with nearby protein side chains and backbones. Naturally interacting proteins within the cell can be instantly trapped by photoactivation of the diazirine-containing proteins in the cultured cells. Crosslinked protein complexes can be detected by decreased mobility on SDS-PAGE followed by Western blotting, size exclusion chromatography, sucrose density gradient sedimentation or mass spectrometry. (en)
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  • Un aminoàcid anàleg fotoreactiu és un tipus d'aminoàcid artificial, anàleg als naturals, que pot ésser incorporat a proteïnes i pèptids amb la principal finalitat de detectar interaccions entre diverses proteïnes. Uns exemples d'aquests aminoàcids fotoreactius són: la fotometionina -amb fórmula molecular C6H11N3O2- que deriva de la metionina (C5H11NO2S), o la fotoleucina (C5H9N3O2) que deriva de la leucina (C6H13NO2). (ca)
  • Photo-reactive amino acid analogs are artificial analogs of natural amino acids that can be used for crosslinking of protein complexes. Photo-reactive amino acid analogs may be incorporated into proteins and peptides in vivo or in vitro. Photo-reactive amino acid analogs in common use are photoreactive diazirine analogs to leucine and methionine, and para-benzoylphenylalanine. Upon exposure to ultraviolet light, they are activated and covalently bind to interacting proteins that are within a few angstroms of the photo-reactive amino acid analog. (en)
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  • Aminoàcid anàleg fotoreactiu (ca)
  • Photo-reactive amino acid analog (en)
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