Team:Fatih Turkey/Reflectin

           Many cephalopods(squid, cuttlefish, octopus)  exhibit remarkable dermal  iridescence (more than meets the eye), a component of their complex,dynamic camouflage and communication. In the species Euprymna scolopes, the light organ iridescence is static and is due to reflectin protein-based platelets assembled into lamellar thin-film reflectors called iridosomes, contained within iridescent cells called iridocytes.

         Reflectin  hierarchical protein assembly is necessity for the responsive, tunable optical  function of iridosome cells. This protein assembly can be triggered by chemical  stimulation and that assembly can be reversible and fine-tuned.

                  Namely reflectin proteins were  identified as the major biomaterial component of iridosomes. The RA1 gene in The Rainbow Graveyard Project was artificially synthesized into  pBluescript to transform into E. coli. In addition, Kramer  et al. (2007) isolated reflectin proteins exhibit unusual solubility and selfassociation properties.

          Self-association of RA1 is likely to stem from a combination of  electrostatic and weak aromatic interactions. Reflectins are, indeed, characterized by their high content of polar aromatic residues and arginines, and several lines of evidence suggest that they are intrinsically unstructured, with no likely transmembrane, alphahelix or beta-sheet regions. E. scolopes reflectins also have high arginine, hydrophobic aromatic content and ‘methionine-rich membrane-associated proteins’. It’s demonstrated that they have a high affinity for assembly with microsomal membranes.
           The mechanism of reflectance is the same as that of coloured soap bubbles. If the soap film (or  multilayer plate) is very thin, shorter wavelengths are reflected, e.g. blue light; if it is thicker, longer wavelengths, such as yellow and red, are reflected.
The reflectin proteins that apparently change their conformation or assembly to reversibly create the photonic structure.

Reflector, formed of a highly organized multilayer of collagen rods that reflect specific wavelengths

            Multilayer reflector that appears red at near-normal viewing angles will appear first yellow, then green and blue at increasingly oblique angles.

           This dynamic reflection occur by altering  platelet and 
           inter-platelet thicknesses in the  multilayer reflector and/or altering 
           the overall effective refractive index of the  intra-platelet material. This allows the entire visible spectrum to be  reflected from a single 
           platelet stack. 
           The  increase in film thickness resulted in detectable redshift 
           of the visible spectra and dominated any effect of decreasing 
           refractive index owing to water sorption,  which would have caused 
           a blue-shift in the spectrum 
           Kramer et al.  (2007) have performed Micro-dialysis  of reflectin 1a into various buffers, which resulted 
           in two general types of aggregative  structures. Optically clear 
           bulk precipitation was seen in non-reducing  conditions 
           and filamentous protein structures were  observed in reducing 
           conditions  controlled through the addition of a 10:1 
           ratio  of reduced to oxidized glutathione 
           After several weeks at 
           4 ◦C, the  filamentous structures formed a webbed structure that 
           resulted in the supramolecular assembly of thin ribbons 

            In our project, we use reflectin 1a for detecting survival  of E.coli. Both of our bacteria (E.coli and B.subtilis) are expected to synthesize reflectin. When the biofilm containing anti-LPS factor (LALF) from B.subtilis, it will bind to LPS on Gram (-) bacteria’s cell wall (E.coli in our project) and so Gram (-) bacteria will not grow and survive in our biofilm surface. Around the binding, reflectin thickness is going to decrease and change the surface colours because of E.coli’s death. So if the colour changes from red to blue, we can say there was a contact between E.coli and B.subtilis via anti-LPS factor.
You can see <a href="https://2011.igem.org/Team:Fatih_Turkey/Anti_LPS">https://2011.igem.org/Team:Fatih_Turkey/Anti_LPS</a>  for more information about LALF.

REFENCES

1)nature materials VOL 6 JULY 2007
(Review.The self-organizing properties of squid reflectin protein, Ryan M. Kramer et al.)
Published online: 3 June 2007; doi:10.1038/nmat1930

2)J. R. Soc. Interface (2009) 6, S149–S163
doi:10.1098/rsif.2008.0366.focus
Published online 15 December 2008
(Review. Structural coloration in cephalopods L. M. Ma¨thger et al. S155)

3)Review. Structural coloration in cephalopods L. M. Ma¨thger et al. S161

4)DOI: 10.1126/science.1091288
Science 303, 235 (2004);
Wendy J. Crookes, et al.(www.sciencemag.org SCIENCE VOL303 9 JANUARY 2004)

5)A.R. Tao et al. / Biomaterials 31 (2010) 793–801

6)J. R. Soc. Interface (2010) 7, 549–560
doi:10.1098/rsif.2009.0299
Published online 23 September 2009(Reflectin phosphorylation M. Izumi et al.)