The Chemistry of

Bioluminescence:

Shedding Light on Cancer Research

Sheila MacIntyre

Cohort 7

Bioluminescent organisms have

been lighting up the seas and

summer nights for centuries and

recently have been shedding light

on cancer research.

Chemiluminescence

•Production of light as the result of a chemical

reaction, during which chemical energy is

converted to light energy

•Bioluminescence

–The production and emission of light by a living

organism (Hastings, 2006)

•Bioluminescent reaction is an extremely efficient

“cold”reaction

–heat is not generated in the production of light

Bioluminescent Organisms

•17 phyla

•700 genera

•70% of marine

organisms are

bioluminescent (Campbell,

2003)

(Haddock, 2008)

MarineEnvironment

http://www.seasky.org/deep-sea/ocean-layers.html#Zone3

Evolution of Bioluminescence

•Why would organisms living >1000 m

need light to survive?

–Camouflage

–Attraction

–Repulsion

–Communication

(Haddock, 2005; Rees, 1998)

Camouflage

•Counterillumination

–Squid

–Fish

–Sharks

•Red pigmented

–Red = Black

(Haddock, 2008)

Camouflage

(Haddock, 2008)

Attraction

fogonazos.blogspot.com/.../glowworm

-heaven.html

Repulsion

(Haddock, 2008)

Communication

•Quorum Sensing

–Bacteria release

autoinducers to coordinate

gene expression according

to local density (Bassler,

2007)

•Photophores

–Species

–Gender

–Territory

–Mating status

(Haddock, 2008)

Evolution of Bioluminescence

•May have served as an antioxidant to

protect organism from oxidative stress

(Hastings, 1983)

•Luciferin is found in peroxisomes of light

producing cells

•Photocytes contain many tracheal

terminals and mitochondria

•Inhibits acellular lipid peroxidation

Bioluminescence Emissions

(Thompson, 2008)

Light Production of Terrestrial

Organisms

(Viviani, 2008)

Production of Light

•Substrate

–Luciferin protein

•Oxidizing Agent

–O2

–an oxygen derivative

–photoprotein

•Enzyme

–Luciferase –a protein catalyst

Enzyme Catalyzed Reaction

Importanceof Luciferin

•Blue photons λ=470 nm

•Organisms need to consume 255 kJ mol-1 energy, E = h

c/λ

•ATP hydrolysis = 30 kJ mol-1

•Luciferin provides a peroxide bond which will provide the energy

through electron transfer by breaking the covalent bond to produce

radical intermediates (Lee, 2002)

Luciferin Molecules

General Luciferin Reaction

Luciferin/luciferase Reaction:

Luciferin + ATP �

luciferyl adenylate + PPi

Luciferyl adenylate + O

2�oxyluciferin* + AMP + CO2+ h

v

Photoprotein Reaction:

Green Fluorescent Protein

Photoprotein + Ca2+�Protein-coelenteramide* + CO2+ h

v

hv= 460-490 nm

(Lee, 2008)

Coelenterazine

inside the hydrophobic GFP

•Calcium binding

protein

–Calmodulin or troponin C

•donates the peroxide

to excite the molecule

Coleoptera Luciferin

Benzothiozole

(Branchini, 2006)

Firefly Luciferase

Benzothiozole

•Environmental influences

–Carboxyl or amine substituted group in solvent

–Enzyme assisted Keto-enol tautomerism

–pH 6.0 = λmax 615 nm keto form shifts

–pH 8 = λmax 560 nm enolate dianion form shifts

–Stereochemistry

(Branchini, 2008)

Bacterial

Reduced flavin and long chain aldehyde

(Meighen, 2008)

“Milky Sea”in the Indian Ocean

Vib

rio h

arv

eyi

Dinoflagellate Luciferin

Tetrapyrrole

λmax = 475 nm

N H

(Z)

O

HN

(Z)

H N(Z

)

NH

O

C

(E)

NaO2C

OO

Bioluminescence Imaging

•Infectious diseases in animal models

•Real time monitoring of progression of

infection

•Pathogens or mice are genetically

encoded with the luciferin gene from

bacteria, insects or the sea pansy

•Light output is related to analyte

concentration

Equipment

•Sample chamber

•Light detector

–Photodiodes and

photomultiplier tubes

in commercial

luminometers

•Signal processing

apparatus

Adding the luciferin gene

Transgenic Mice

Bioluminescent Transgenic Mice

Genetically Modified Organisms

Luc Gene

(Moriyama, 2004)

VEGFR2-luc-KI mouse developing a subcutaneous

A549 xenograft tumor

Conclusion

•Cancer treatments

•New Antibiotic Strains

•Bacteria Communication

•Genetic Markers

•Infectious Diseases

•Biological Tags

•Biological Warfare

•Marine Ecosystems

•Novelty Items

References

1.Haddock, S.H.D., C.W. Dunn, P.R. Pugh, C.E. Schnitzler.(2005) Bioluminescent

and red fluorescent lures in a deep-sea siphonophore.

Scie

nce. Vol 309. p262.

2.Birdsey, R.(2008)Bioluminescence, Light of Life.Retrieved April 2, 2008.from

Website,http://www.nswseakayaker.asn.au/mag/46/bioluminescence.html

3.Bassler, B.(2007)Cell to Cell Communication in Bacteria.Retrieved April 2, 2008,

from Website, http://www.hhmi.org/research/investigators/bassler.html

4.Rees, J., B. De Wergifosse, O. Noiset, M. Dubuisson, B. Janssens, E.M.

Thompson.(1998)The origins of marine bioluminescence:Turning oxygen defense

mechanisms into deep-sea communication tools.

J. E

xp. B

iol. 201: 1211-12

5.Hastings, J.W. and Morin, J.G. I

n G

reen

Flu

ore

scent P

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in:

Pro

pert

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Applic

ations, and P

roto

cols

, S

econd E

ditio

n; Chalfie, M. and Kain, S.R.; Second

Edition; John Wiley and Sons, Inc. New York, NY, 2006: pp 15-38.

6.Case, J.F. (2008) Overview of the last twenty years in marine bioluminescence

research. Retrieved from Website, March 19, 2008.

www.lifesci.ucsb.edu/~biolum/sdworkshop/caseabs.html

7.Lee, J. and Vysotski, E.S. Structure and spectra in bioluminescence. Retrieved from

Website, Jul 25, 2008. http://www.photobiology.info/develop/AdvModsBiolumLee.asp

8.Hastings, J.W. (1983) Biological diversity, chemical mechanisms, and the

evolutionary origins of bioluminescent systems. J

. M

ol. E

vol. 19:309-321.

References

9. Thompson, C.M. B

asic

Bio

lum

inesce

nce

.Retrieved from Website, July 27,

2008. http://www.photobiology.info/develop/biolumin.asp

10. The Deep Sea Anglerfish. Retrieved from Website July 27, 2008,

http://www.starfish.ch/frogfish/Deepsea-Anglerfish.html

11.Scripps Institution of Oceanography. (2008)A history of bioluminescence

according to E.N. Harvey. Retrieved from Website March 19, 2008,

http://siobiolum.ucsd.edu/Biolum_Harvey.html15.

12.Moriyama, E.H., S.K. Bisland, L. Lilge, B.C.

Wilson.(2004)Bioluminescence Imaging of the Response of Rat

Gliosarcomato ALA-PpIX-mediated Photodynamic Therapy.Retrieved

from Website April 20, 2008,

http://findarticles.com/p/articles/mi_qa3931/is_200409/ai_n9426918/pg_1

13.Iyer, M., M. Sato, M. Johnson, S.S. Gambhir, L.Wu.(2005)Application of

Molecular Imaging in Cancer Gene Therapy.Current Gene Therapy, 5:

607-618.

14. Meighen, E.ChenLin, L.Y. 2008. The biochemistry and molecule biology

of bacterial bioluminescence. Retrieved from Website July 30, 2008,

http://www.photobiology.info/develop/AdvModsBiolumMeighen.asp.