HOMEWORK#3

Electronic Journal / Virtual Library

• due on 4/15

• The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is rapidly becoming an important reporter molecule for monitoring gene expression and protein localization in vivo, in situ and in real time. GFP emits bright green light (lambda max = 509 nm) when excited with UV or blue light (l max = 395 nm, minor peak at 470 nm). The fluorescence excitation and emission spectra of GFP are similar to those of fluorescein, and the conditions used to visualize this fluorophore are also suitable for GFP. Unlike other bioluminescent reporters, the chromophore in GFP is intrinsic to the primary structure of the protein, and GFP fluorescence does not require a substrate or cofactor. GFP fluorescence is stable, species-independent and can be monitored non-invasively in living cells and, in the case of transparent organisms, whole animals.
• How many references can you find about green fluorescent protein (GFP) within five years ?
• How many papers were done by P. Kitts ?
• How many of these references are related to structure ?
• Can you find a paper describe its crystal structure ? Give its abstract on your homepage.

 Answers:

1.

Total:614

2.

There are 21 parpers were done by P.Kitts

3.

There are 76 references are related to structure within 5 years.

There are 82 references are related to structuer within no limit years.

4.

I can find 5 references about GFP crystal structure :

Li X, et al. [See Related Articles] Deletions of the Aequorea victoria green fluorescent protein define the minimal domain required for fluorescence. J Biol Chem. 1997 Nov 7; 272(45): 28545-28549. PMID: 9353317; UI: 98019228.

Wachter RM, et al. [See Related Articles] Crystal structure and photodynamic behavior of the blue emission variant Y66H/Y145F of green fluorescent protein. Biochemistry. 1997 Aug 12; 36(32): 9759-9765. PMID: 9245407; UI: 97392641.

Brejc K, et al. [See Related Articles] Structural basis for dual excitation and photoisomerization of the Aequorea victoria green fluorescent protein. Proc Natl Acad Sci U S A. 1997 Mar 18; 94(6): 2306-2311. PMID: 9122190; UI: 97225948.

Ormo M, et al. [See Related Articles] Crystal structure of the Aequorea victoria green fluorescent protein. Science. 1996 Sep 6; 273(5280): 1392-1395. PMID: 8703075; UI: 96355665.

Perozzo MA, et al. [See Related Articles] X-ray diffraction and time-resolved fluorescence analyses of Aequorea green fluorescent protein crystals. J Biol Chem. 1988 Jun 5; 263(16): 7713-7716. PMID: 2897362; UI: 88227972.

And one of the abstract is:

Science 1996 Sep 6;273(5280):1392-1395

Crystal structure of the Aequorea victoria green fluorescent protein.

Ormo M, Cubitt AB, Kallio K, Gross LA, Tsien RY, Remington SJ

Institute of Molecular Biology and Department of Physics, University of Oregon, Eugene, OR 97403-1226, USA.

The green fluorescent protein (GFP) from the Pacific Northwest jellyfish Aequorea victoria has generated intense interest as a marker for gene expression and localization of gene products. The chromophore, resulting from the spontaneous cyclization and oxidation of the sequence -Ser65 (or Thr65)-Tyr66-Gly67-, requires the native protein fold for both formation and fluorescence emission. The structure of Thr65 GFP has been determined at 1.9 angstrom resolution. The protein fold consists of an 11-stranded beta barrel with a coaxial helix, with the chromophore forming from the central helix. Directed mutagenesis of one residue adjacent to the chromophore, Thr203, to Tyr or His results in significantly red-shifted excitation and emission maxima.