TurboGFP: Properties, Sequence, MW, origin

Everything you need to know about TurboGFP: properties and applications

TurboGFP is a bright green fluorescent protein used to study protein function, localization and dynamics in cells. Nanobody based research tools allow reproducible biochemical analysis including mass spectrometry and enzyme activity measurements of TurboGFP fusion proteins.

1. What are the differences between TurboGFP and GFP

Origin of turboGFP

TurboGFP is a bright dimeric green fluorescent protein derived from CopGFP of the copepod Pontellina plumata. CopGFP has been first published in 2004 (Shagin et al., 2004)

Origin of GFP

GFP has been originally isolated from jellyfish Aequorea Victoria. There are many GFP variants available such as EGFP, GFP S65T, YFP, CFP, mPhluorin, etc. These fluorescent protein derivatives differ in both functional and spectral properties, which have been subject to optimization. The first major improvement of GFP was a single substitution (S65T), which increased fluorescence intensity & stability and shifted the main excitation peak to 488 nm (Heim et al., 1995). EGFP is a further developed version of GFP, which allows the practical use of GFPs in a variety of different organisms and cells.

2. What are the physical-chemical properties of TurboGFP and EGFP?

TurboGFP vs. EGFP
	TurboGFP	EGFP
Spectrum, Wavelength
-   Excitation max	482 nm	488 nm
-   Emission max	502 nm	509 nm
Structure	dimer	monomer
Size, Molecular Weight	2 x 26 kDa	26.9 kDa
Length	232 aa	239 aa

3. What is the sequence similarity between TurboGFP and EGFP: Sequence comparison

Copepod TurboGFP is evolutionarily distant from jellyfish-derived fluorescent proteins such as EGFP: Turbo-GFP shares only ~20 % sequence identity with the commonly used GFP variants.

Please note: Most anti-GFP antibodies do not bind to TurboGFP, most anti-TurboGFP antibodies do not bind GFP and variants.

Fluorescent Jelly

4. What is the sequence of TurboGFP

For DNA-sequence, protein-sequence and vector plasmids containing TurboGFP, please contact Evrogen: http://evrogen.com/products/products-index-alph.shtml,   customer-support@evrogen.com

5. TurboGFP variants

Other variants of TurboGFP are maxGFP and maxFP-Green.

6. What reliable tools for analyzing TurboGFP-tagged proteins are available?

Currently, there are only few tools for cell biological and biochemical analysis of TurboGFP-tagged proteins commercially available. For

• Immunoprecipitation (IP) of Turbo-GFP fusion proteins

Anti-TurboGFP Nanobody coupled to beads for IP & Co-immunoprecipitation (Co-IP), i.e. for mass spec (MS) analysis: TurboGFP-Trap (ChromoTek)

The TurboGFP-Trap works also with copGFP, maxGFP and maxFP-Green.

• Western blot (WB), Immunoblotting and ICC

Rabbit polyclonal anti-TurboGFP(d) antibody (Evrogen)

• ELISA

Purified anti-TurboGFP Nanobody: TurboGFP- Binding Protein (ChromoTek)

7. What is best anti-TurboGFP antibody for immunoprecipitation?

TurboGFP-Trap, an anti-TurboGFP Nanobody coupled to agarose or magnetic agarose beads, is advantageous over conventional anti-turboGFP antibodies in IP:

• High binding affinity (K_D = 0.5 nM)
• No contaminating antibody heavy and light chains
• Low background
• Stable under harsh washing conditions

Links:

http://evrogen.com/products/TurboGFP/TurboGFP_Detailed_description.shtml

Shagin DA, Barsova EV, Yanushevich YG, Fradkov AF, Lukyanov KA, Labas YA, Semenova TN, Ugalde JA, Meyers A, Nunez JM, Widder EA, Lukyanov SA, Matz MV. GFP-like proteins as ubiquitous metazoan superfamily: evolution of functional features and structural complexity, Mol Biol Evol. 2004 May;21(5):841-50. Epub 2004 Feb 12.

Heim R, Cubitt A, Tsien R. Improved green fluorescence, Nature.1995, Feb; 373 (6516): 663–4.