Protein photochemical cross-linking

Other clues to the self-association of recombinant resilin in solution, and thus a degree of defined stmcture, include the propensity of the monomer proteins to covalently cross-link very rapidly through dityrosine side chains using a mthenium-based photochemical method [29]. Proteins which do not naturally self-associate do not form biomaterials when exposed to the Ru(ll)-based photochemical procedure (Elvin, C.E. and Brownlee, A.G., personal communication). Furthermore, Kodadek and colleagues showed that only intimately associated proteins are cross-linked via this zero-A photochemistry procedure [45]. 

Fig. 12 Illustration outlining MIP film fabrication. The C-terminus nonapeptide epitope is attached through a tether to a glass or oxidized silicon surface by the N-terminal amino acid of the peptide. Monomers are photochemically cross-linked while remaining in contact with the peptide modified surface. Following polymerization, the glass substrate is removed. The protein can now bind to the MIP via its C-terminus nonapeptide epitope. Modified from [114]...

In the V-ATPase, the presence of at least two peripheral stators has been reported (Boekema et al., 1999 Domgall et al., 2002 Wilkens et al., 1999, 2004). Candidates for the stator proteins in the V-ATPase are subunits E and G, and support for this assignment comes from photochemical cross-linking studies that place these subunits on the outside of the Vi domain for much of its length (Arata et al., 2002a,b). Another subunit that probably plays the role of a stator in the V-ATPase is the a-subunit of the Vo- The Vo a-subunit has a large cytoplasmic domain, and it has been shown that this domain interacts with the A-subunit of the Vi (Landolt-Marticorena et al, 2000). 

Bitan G, Teplow DB. Rapid photochemical cross-linking-a new tool for studies of metastable, amyloidogenic protein assemblies. Acc. Chem. Res. 2004 37 357-364. 

P. E. Nielsen, J. B. Hansen and O. Buchardt, Photochemical cross-linking of protein and DNA in chromatin. 1. Synthesis and application of a photosensitive cleavable derivative of 9-aminoacridine with 2 photoprobes connected through a disulfide-containing linker, Biochem. J., 1984, 223, 519-526. 

Cysteine had also been found to add photochemically to poly U, poly C, and DNA, thus adding credence to Smith and Aplin s postulate that the formation of a heterodimer between a pyrimidine and a sulfur (or hydroxy) amino acid may constitute a mechanism for the photochemical cross-linking of DNA and protein in vivo (Smith, 1962, 1964 Smith et al, 1966). Infrared data suggested that the COOH and amino groups of the cysteine residue were free, implying that the linkage to the uracil skeleton was through the sulfur bond. 

Reactive noncanonical amino adds have also been integrated into elastin-like protein polymers to enhance their application as biomaterials. The Tinell laboratory has used the photoreactive amino add p-azidophenylalanine to pattern elastin-like protein polymers on surfaces. " Zhang et d. used p-azidophenylalanine to photochemically cross-link an elastin-leudne zipper protein to a glass surface. A target protein linked to a second leudne zipper domain can then be captured from solution and patterned on the surface by dimerization of the leucine zipper domains. Carrico et al. used similar p-azidophenylalanine-fiinctionalized elastin protein polymers... 

Steen, H. and Jensen, O.N. (2002) Analysis of protein-nucleic acid interactions by photochemical cross-linking and mass spectrometry. Mass Spectrom. Rev., 21 (3), 163-182. 

Another important photoaddition is that of cysteine (in protein) to thymine (in DNA), which can lead to photochemical cross-linking of DNA to protein as illustrated below. 

Blattler, W.A., Kuenzi, B.S., Lambert, J.M., and Senter, P.D. (1985b) New heterobifunctional protein cross-linking reagents and their use in the preparation of antibody-toxin conjugates. Photochem. Photobiol. 42, 231. 

Gorman, J.J., and Folk, J.E. (1980) Transglutaminase amine substrates for photochemical labeling and cleavable cross-linking of proteins./. Biol. Chem. 255, 1175. 

Rabow LE, Stubbe J, Kozarich JW (1990) Identification and quantitation of the lesion accompanying base release in bleomycin-mediated DNA degradation. J Am Chem Soc 112 3196-3203 Raleigh JA, Blackburn BJ (1978) Substrate conformation in 5 -AMP-utilizing enzymes 8,5 -cycloadenosine 5 -monophosphate. Biochem Biophys Res Commun 83 1061-1066 Ramakrishnan N, Clay ME, Xue L-Y, Evans El El, Rodriguez-Antunez A, Oleinick NL (1988) Induction of DNA-protein cross-links in Chinese hamster cells by photodynamic action of chloroaluminium phthalocyanine and visible light. Photochem Photobiol 48 297-303 Ramakrishnan N, Chiu S-M, Oleinick NL (2003) Yield of DNA-protein cross-links in y-irradiated Chinese hamster cells. Cancer Res 47 2032-2035... 

K. Ishihara, N. Hamada, S. Kato and I. Shinohara, Photoresponse of the release behavior of an organic compound by a azoaromatic polymer device, J. Polym. Sci., Polym. Chem. Ed., 1984, 22, 881 K. Ishihara and I. Shinohara, Photoinduced permeation control of protein using amphiphilic azoaromatic polymer membrane, J. Polym. Sci., Polym. Lett. Ed., 1984, 22, 515-518 J. Matejka, M. Ilavsky, K. Dusek and O. Wichterle, Photochemical effects in cross-linked photochromic polymer, Polymer, 1981, 22, 1511. 

Photochemistry of proteins is derived from diose of its amino acid residues. In addition, important photochemical processes in proteins involve the splitting of the disulfide bridges. Furthermore, DNA-protein cross-linking involves the addition of thymine in DNA with cystine in protein. This addition process has been discussed above. In the case of DNA, UV irradiation can also lead to the breaking of either one or both DNA strand and infra- and inter-molecular DNA cross-linking. The photochemistry of RNA is very similar to that of DNA. 

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