Phycobiliprotein: Difference between revisions

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Phycobiliproteins are water-soluble proteins present in cyanobacteria and certain algae (rhodophytes, cryptomonads, glaucocystophytes) that capture light energy, which is then passed on to chlorophylls during photosynthesis. Phycobiliproteins are formed of a complex between proteins and covalently bound phycobilins that act as chromophores (the light-capturing part). They are most important constituents of the phycobilisomes.

The major phycobiliproteins are :

Phycobiliprotein	MW (Da)	Ex (nm) / Em (nm)	Quantum yield	Molar Absorbtion	Comment
R-Phycoerythrin (R-PE)	240 000	498.546.566nm / 576nm	0,84	1.53 10⁶	Can be excited by Kr/Ar laser
B-Phycoerythrin (B-PE)	240 000	546.566nm / 576nm	0,98	(545nm) 2.4 10⁶ (563nm) 2.33 10⁶
C-Phycocyanin (CPC)	232 000	620nm / 642nm	0,81	1.54 10⁶	Accepts the fluorescence for R-PE; Its red fluorescence can be transmitted to AllophycoCyanine
Allophycocyanin (APC)	105 000	651nm / 662nm	0,68	7.3 10⁵	Excited by He/Ne laser; double labeling with Sulfo-Rhodamine 101 or any other equivalent fluorochrome.
↑ = FluoProbes PhycoBiliProteins datas

Characteristics and Applications in Biotechnologies

Phycobiliproteins elicit great fluorescent properties compared to small organic fluorophores, especially when high sensitivity or multicolor detection is required :

Broad and high absorption of light suits many light sources
Very intense emission of light: 10-20 times brighter than small organic fluorophores
Relative large Stokes shift gives low background, and allows multicolor detections.
Excitation and emission spectra do not overlap compared to conventional organic dyes.
Can be used in tandem (simultaneous use by FRET) with conventional chromophores (i.e. PE and FITC, or APC and SR101 with the same light source).
Fluorescence retention period is longer.
Very high water solubility

As a result, phycobiliproteins allow very high detection sensitivity, and can be used in various fluorescence based techniques (Fluorimetry in microplate, Flow Cytometry, FISH, two or multicolor detections…).

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@@ Line 6: / Line 6: @@
 |-
 ! '''Phycobiliprotein'''
-! MW (Da)
+! MW ([[atomic mass unit|Da]])
 ! Ex (nm) / Em (nm)
 ! Quantum yield
@@ Line 12: / Line 12: @@
 ! Comment
 |-
-| '''R-[[Phycoerythrin]]'''
+| '''R-[[Phycoerythrin]] (R-PE)'''
 | 240 000
-| 498.546.566nm/576nm
+| 498.546.566nm / 576nm
 | 0,84
 | 1.53 10<sup>6</sup>
 | Can be excited by Kr/Ar laser
 |-
-| '''B-[[Phycoerythrin]]'''
+| '''B-[[Phycoerythrin]] (B-PE)'''
 | 240 000
-| 546.566nm/576nm
+| 546.566nm / 576nm
 | 0,98
 | (545nm) 2.4 10<sup>6</sup>
 (563nm) 2.33 10<sup>6</sup>
 |-
-| '''C-[[Phycocyanin]]'''
+| '''C-[[Phycocyanin]] (CPC)'''
 | 232 000
-| 620nm/642nm
+| 620nm / 642nm
 | 0,81
 | 1.54 10<sup>6</sup>
 | Accepts the fluorescence for R-PE; Its red fluorescence can be transmitted to AllophycoCyanine
 |-
-| '''[[Allophycocyanin]]'''
+| '''[[Allophycocyanin]] (APC)'''
 | 105 000
-| 651nm/662nm
+| 651nm / 662nm
 | 0,68
 | 7.3 10<sup>5</sup>
+| Excited by He/Ne laser; double labeling with Sulfo-Rhodamine 101 or any other equivalent fluorochrome.
 |-
 | colspan=6 rowspan=1 | ↑ = [[FluoProbes]] [http://www.interchim.fr/ft/2/28310A.pdf| PhycoBiliProteins datas]
 |-
 |}
+=== Characteristics and Applications in Biotechnologies ===
+Phycobiliproteins elicit great fluorescent properties compared to small organic fluorophores, especially when high sensitivity or multicolor detection is required :
+* Broad and high absorption of light suits many light sources
+* Very intense emission of light: 10-20 times brighter than small organic fluorophores
+* Relative large Stokes shift gives low background, and allows multicolor detections.
+* Excitation and emission spectra do not overlap compared to conventional organic dyes.
+* Can be used in tandem (simultaneous use by [[Förster_resonance_energy_transfer|FRET]]) with conventional chromophores (i.e. PE and FITC, or APC and SR101 with the same light source).
+* Fluorescence retention period is longer.
+* Very high water solubility
+As a result, phycobiliproteins allow very high detection sensitivity, and can be used in various fluorescence based techniques (Fluorimetry in microplate, Flow Cytometry, FISH, two or multicolor detections…).
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