Download Poster

St. Dominic SMART Team: Elana Baltrusaitis, Allyson Bigelow, Rachel Brielmaier, Pamela Burbach, Jake Dowler, Johnny Fuller, Caroline Hildebrand, Teagan Jessup, Molly Jordan, Josh Kramer,
Jenna Lieungh, Alex Mikhailov, Pat O’Grady, Andrew Pelto, Quin Rowen, Rachelle Schmude, Bobby Schultz, Katherine Seubert, Alex Sherman, Parker Sniatynski, Alex Venuti, Erin Verdeyen, and Molly Wetzel
Teacher Advisor Donna LaFlamme
Mentors: Nathan Duncan and Françoise Van den Bergh, Ph.D., Medical College of Wisconsin
Luciferase is the generic name for an enzyme responsible for bioluminescence reactions and is commonly associated with fireflies. It is also found in many other organisms
including bacteria, fungi, anemones, and dinoflagellates. Since the gene for the North American firefly (Photinus pyralis) luciferase was cloned in 1985, scientists have been
genetically engineering the gene into living cells. The luciferase reaction is now widely used in scientific research to study protein production in cells, to analyze gene promoter
activity, to study stem cell function in vivo, and in cancer studies, to trace the metastasis of cancer cells in living test animals. The scientific study of the luciferase enzymes
themselves is also continuing. In recent research, single amino acid mutations to the active site cause the emission of different colored light in a predictable way. The uses of and
improvements in bioluminescent imaging are increasing exponentially in cell biology, molecular biology, and in medical research.
LUCIFERASE FACTS TO KNOW
LUCIFERASE IN RESEARCH
• Luciferase is an enzyme found in the lanterns of fireflies in cells called
photocytes.
• Its function is to catalyze a bioluminescence reaction in which the substrate
luciferin is oxidized to oxyluciferin.
• Bioluminescence is the production of light by a chemical reaction within
living organisms.
• Fireflies use bioluminescence for communicating, attracting mates, luring in
prey, and self-defense.
Firefly Luciferase (Luciola cruciata)
Physical Model based on PDB File: 2D1R
[1]
Fat Cell Growth Over 12 Weeks
ACTIVE SITE OF FIREFLY LUCIFERASE
(Luciola cruciata)
Japanese Firefly (Luciola cruciata)
Scientists discovered a fat progenitor cell
with the help of luciferase. The progenitor
cells were injected into mice genetically
unable to produce fat. The luciferase gene
turned on in mature fat cells and allowed the
researchers to track the development of the
fat cells over time in the living animal. [2]
http://2.bp.blogspot.com/_VVjUm4hzpvo/SSPbxn1OnJI/AA
AAAAAABd8/UnJeVFSM9RI/s400/firefly.jpg
IVIS® imaging system which is used
to pick up the faint light given off
by the oxyluciferin in the cells
expressing luciferase.
ACTIVE SITE OF WILD TYPE AND MUTANT S286N LUCIFERASE
FIREFLY BIOLUMINESCENCE REACTION
Ile 288
DLSA
RasMol Image based on PDB File: 2D1R
[1]
Step 1: Luciferin and magnesium-ATP react to form the luciferyl-AMP
intermediate.
Step 2: Luciferin-AMP reacts with O2 to form excited oxyluciferin which
emits a photon of light as it returns to its ground energy state.
1885- Raphael DuBois ground up beetle
lanterns in hot and cold water obtaining
no luminescence in the hot water. He named luciferin and
luciferase. Hot water destroyed luciferase. [3]
1955-Luciferase was purified as
a protein by Green and McElroy
The luciferase active site provides a hydrophobic pocket for
producing the electronically unstable oxyluciferin. Single
amino acid mutations of Ile 288 to Val288 and to Ala288 and
of Ser 286 to Asn 286 results in the emission of lower energy
light. [1] See below.
Wild Type(yellow) I288V (orange) l288A (reddish-pink) S286A(red)
1947- W.D. McElroy discovered that ATP is an
essential requirement for the bioluminescence
reaction and light production is proportional to ATP. [3]
1959- McElroy and Seliger discovered the
quantum yield of the oxidation of
firefly luciferin to be 90% efficient.[3]
bioluminescencehttp://upload.wikimedia.org/wikipedia/en/2/27/William_David_McElroy.jpg
http://upload.wikimedia.org/wikipedia/commons/f/f7/Luciferase.png
[1]Primary Citation : T. Nakatsu, et. al. Structural basis for the spectral difference in luciferase bioluminescence. Nature, 2006, 440, 372-376
1976- DeLucca shows that the firefly
bioluminescence reaction is a two step
process requiring ATP, O2, and
luciferin.
1967- Determination of picogram amounts
of ATP using luciferin-luciferase enzyme
system by G.E. Lyman and
J.P. DeVincenzo. [4]
[2] M. S. Rodeheffer et. al. Identification of White Adipocyte Progenitor Cells In Vivo. Cell, 2008, 135240-249
[3] H. Fraga, Firefly luminescence: A historical perspective and recent developments. Photochemical and Photobiological Sciences, 2008, 7, 146-158
[4] J. Lee, Bioluminescence Timelines, University of Georgia.
A SMART Team project supported by the National Institutes of Health (NIH)-National Center for Research Resources Science Education Partnership Award (NCCR-SEPA).
Wild-Type
DLSA
S286N
Mutant
Ile 288 is being rotated so it is not packing the intermediate analogue,
DSLA ,tightly in the mutant active site. This changes the color of the
emitted light from green to lower energy red light because excited
oxyluciferin is allowed to lose energy before it emits light. [1]
[1]
1961-The structure and synthesis of
luciferin was determined
by White, Field, McElroy,
and McCapra.[4]
Ile 288
1985-Marlene DeLucca and
colleagues cloned the gene for
firefly luciferase
and expressed the
gene in E. coli bacteria.
1990/91-Mutant luciferases
discovered that produce
different colors of light
http://hongkiat.s3.amazonaws.com/colorfulwp/Rainbow_Ocean__by_Thelma1.j
pg
Present- luciferase/luciferin is
used to locate tumors and monitor the
spread of cancer in test animals
1996- E. Conti, N. Frank,
and P. Brick crystallized
firefly luciferase and
obtained an x-ray structure
at 2 angstroms resolution.
2010-St.Dominic
SMART Team designs
physical model of firefly
luciferase (2d1r)