Synthesis and characterization of a long, rigid photoswitchable cross-linker for promoting peptide and protein conformational change.
Azobenzene-based photoswitchable compounds can be use to photocontrol a variety of biochemical systems. In some cases, their effectiveness may be limited by the size of the conformational change that the switch undergoes. To produce an azobenzene photoswitch that undergoes a large end-to-end distance change upon isomerization, we synthesized 3,3'-diazene-1,2-diylbis{6-[2-sulfonato-4-(chloroacetylamino)phenylethynyl]benzene sulfonic acid} (DDPBA). This long, rigid, water-soluble, thiol-reactive cross-linker undergoes an end-to-end distance change of approximately 13 A upon isomerization. DDPBA was successfully cross-linked to peptides through cysteine side chains. The photoswitch undergoes trans-to-cis photoisomerization maximally when irradiated at 400 nm, although the efficiency of production of the cis isomer is lower than for simpler azobenzenes. Under steady-state illumination conditions, the percentage of cis form produced increases as temperature increases; approximately 56 % cis is obtained at 60 degrees C. Thermal relaxation occurs with a half-life of approximately 75 min at room temperature. When DDPBA was attached to an alpha-helical peptide with two cysteine residues at i and i+14 positions, an increase in helix content was observed after photoirradiation. When cross-linked to another peptide with two cysteine residues spaced at i and i+21 positions, a decrease in helix content after trans-to-cis isomerization was observed. Due to the small percentage of cis form produced under the experimental conditions, the CD signal changes were small. However, the large structural change upon photoisomerization provided by this cross-linker can potentially be used to photoswitch other biochemical systems.
Zhang
Fuzhong
F
Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S3H6, Canada.
Sadovski
Oleg
O
Woolley
G Andrew
GA
D000595:Amino Acid Sequence; D001391:Azo Compounds / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001557:Benzenesulfonates / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D002942:Circular Dichroism; D003432:Cross-Linking Reagents / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015195:Drug Design; D007536:Isomerism; D008027:Light; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010777:Photochemistry; D011487:Protein Conformation / Q000187:drug effects; D017433:Protein Structure, Secondary; D012997:Solvents / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet
C534886:3,3'-diazene-1,2-diylbis(6-(2-sulfonato-4-(chloroacetylamino)phenylethynyl))benzenesulfonic acid; D001391:Azo Compounds; D001557:Benzenesulfonates; D003432:Cross-Linking Reagents; D010455:Peptides; D012997:Solvents