The laboratory of Plant Biochemistry and Photobiology (LPBP), ex-LPPBP, which is directed by C. A. Rebeiz, was a research unit of the Department of Natural Resources and Environmental Sciences
(NRES) within the College of Agricultural Consumer and Environmental Sciences (ACES), University of
Illinois, Urbana Campus. It was founded with a mission of studying the chemistry and biochemistry of the GREENING PROCESS. Since its inception in 1972, LPBP grew into a multidisciplinary research center with several
major discoveries to its credit. In 1983, the laboratory won a remodeling for excellence award of about 1 million dollars from the University
and State of Illinois. The award was used to remodel new research facilities. In July 1986, LPBP moved into about 5000 square feet of state
-of-the art remodeled research space. Then in December 1986, the laboratory received a monetary gift from John P. Trebellas, an Urbana-
Champaign philanthropist, to establish the John P. Trebellas Photobiotechnology Research Endowment aimed at enabling LPBP to become a research
catalyst among various departments within and outside ACES and at "Bettering the Human Condition Through Basic and Agricultural
Photobiotechnological Research". In April, 1991, LPBP moved into the newly constructed 30 million dollar University of Illinois Edward Robert
Madigan Laboratory (ERML) where LPBP functions in close proximity with plant and animal molecular biologists, biophysicists, biochemists and
physiologists. In 1996 LPBP received the C. A and C. C. Rebeiz Endowment for Basic Research with matching funds from ACES and NRES.
Proceeds from the various endowments were used to support graduate study within LPBP. In November 1998, professor Donald P. Briskin, a well
known plant biochemist with interest in medicinal plant chemistry and biochemistry, joined forces with LPPBP.
On June 30, 2005, with the retirement of C. A. Rebeiz from the University of Illinois, and the establishment of the Rebeiz Foundation for
Basic Research, the Laboratory of Plant Biochemistry and Photobiology has been converted into the Virtual Laboratory of Plant Biochemistry
and Photobiology (VLPBP). In Addition to its dedication to the promotion of research dealing with the greening process, the VLPBP website will
be expanded to become the voice of the Rebeiz Foundation for Basic Research. The Foundation will be a clearing house for the promotion of
Chloroplast Bioengineering Nationally and Internationally. Among other things, the Foundation will sponsor national and international
research on Chloroplast Bioengineering, will feature papers of the month and papers of the year to promote the best research in chloroplast
bioengineering, and will deliver annual prizes for the best papers in the field. It will also undertake to match applicants with available
positions in Chloroplast Bioengineering.
At the present the Governing Board of the Foundation consists of:
Constantin A. Rebeiz (Director Rebeiz Foundation for Basic Research)
Christopher Benning (Michigan State University)
Hans Bohnert (University of Illinois)
Henry Daniell (Central Florida University)
Natalia Dudareva (Purdue University)
Kenneth Hoober (SolaraPharma)
Archibald Portis (University of Illinois)
Baishnab Tripathy (Nehru University)
Julian Whitelegge (University of California at Los Angeles)
Carole Rebeiz, (Secretary, Rebeiz Foundation for Basic Research)
First International Symposium On Chloroplast Bioengineering: University of Illinois, Allerton
Conference Center, May 2-May 7, 2005. Click here to enter the website and register
.
The First International Symposium on Chloroplast Bioengineering was held in Urbana on May 2-7, 2005. The proceedings of the Symposium will be
published in 2007.
A description of a Post-acquisition software for quantitative calculation of the porphyrin and phorbin metabolic pools of the heme and
chlorophyll biosynthetic pathways by spectrofluorometry is described HERE.
To Calculate a porphyrin concentration, and if you are a beginner, click on the HERE link. If you are an experienced user click on
the "Porphyrin Analytical Tools" link and
follow the described step-by-step instructions. When the Java Security Warning Window appears, click on the Grant This Session
button. In order to perform quantitative calculations with the Java routine, it is mandatory to download and install the Java (TM) 2 Runtime
Environment Standard Edition file named: J2re-1_4_0_01-windows-i586. This self-extractable and -installable file can be downloaded from the
following website: http://java.sun.com, by clicking the "Download J2seTMv1.4 Now button.
Note: This program was developped by Professor Daniel G. Saab of Case Western University.
The program is meant to handle spectral data generated on fully corrected SLM DW2 spectrofluorometers with Data.dat file structure. Later on a
similar program will be added to handle spectral data acquired on fully corrected SPEX spectrofluorometers with Data.Spc file format. It
should be kept in mind that the equations were compiled with photomultiplier sentivities of LPBP SLM Spectrofluorometers. Individuals who wish
to obtain accurate computations should construct calibration curves that relate known porphyrin concentrations to the web-calculated
concentrations. In this manner the user would have a correction factor for every listed equation that relates their instrument sentivity to
LPBP's.
- Copy the Data.dat file containing the spectral data anywhere to the hard drive or keep it on a removable diskette.
- Browse to the location of the Data.dat file and doudle click on it. Click on the "ClickHereToLoadData" button to load the data on
the computer.
- Select a porphyrin equation such as Proto....E400, E440 from the dropdown menu.
- Enter the spectral location (a 1-83 number) for the E400 spectrum, i.e.for the emission spectrum elicited by excitation at 400 nm.
- Enter the spectral location (a 1-83 number) for the E440 spectrum, i.e.for the emission spectrum elicited by excitation at 440 nm.
- Click on "ClickHereToDisplayResults" The results i. e. the Proto concentration in pmole/ml +/- standard deviation is
displayed in the display window.
- The "Total_Q" entry is meant for the MV, DV equations. An entry of 100 will yield the percent of the MV and DV components for a specific
MV + DV mixture of the tetrapyrrole. An entry of the total MV + DV tetapyrole content will yield the actual amounts of MV and DV
tetrapyrrole components in pmoles/ml.
- Stephen O. Duke and C A. Rebeiz (1994). Porphyric Pesticides: Chemistry,Toxicology and Pharmaceutical Applications, S.O. Duke and
C. A. Rebeiz, eds. ACS Symposium Series 559 pp 317.
- Rebeiz, C. A. (1994). Photodynamic Herbicidal Compositions Using Delta-Aminolevulinic Acid. U. S. Patent Ser. No. 5,286,708
pp 38.
- Rebeiz, C. A., Juvik, J. A. and Rebeiz C. C. (1994) Porphyric insecticides. U. S. Patent Ser. No 5300526, pp. 18.
- Rebeiz, C. A. (1994) Photodynamic Defoliants. U. S. Patent Ser. No 5321001, pp. 23.
- Gut, L. J., Lee, K., Juvik, J. A. Rebeiz, C. C. and Rebeiz, C. A. (1994) Porphyric Insecticides 6. Structure Activity Study of
Substituted Pyridyls. Pest. Biochem. Physiol. 50: 1-14.
- Ioannides, I., Fasoula D. A., Robertson, K. R. and Rebeiz, C. A. An evolutionary study of chlorophyll biosynthetic heterogeneity in
green plants.(1994) Biochem. Syst. Ecol. 22:211-220.
- Rebeiz, C. A., Parham, R., Fasoula, D. A., and Ioannides, I. M. (1994) Chlorophyll a Biosynthetic Heterogeneity. In: Biosynthesis of
tetrapyrrole pigments. Ciba Symposium 180. pp 177-193, John Wiley, New York.
- Duke S. O. and Rebeiz, C. A. (1994). Porphyrinogenesis as a tool in pest management. In: Porphyric Pesticides: Chemisry, Toxicology, and
Pharmaceutical Applications (S. O. Duke and C. A. Rebeiz, eds.) ACS Symposium Series 559, pp 1-16
- Rebeiz C. A., Amindari S., Reddy N. K., Nandihalli U. B. Moubarak M. B., and Velu, J. A. (1994) Delta-aminolevulinic Acid-Based
Herbicides . In: Porphyric Pesticides: Chemisry, Toxicology, and Pharmaceutical Applications (S. O. Duke and C. A. Rebeiz, eds.) ACS
Symposium Series 559 pp. 48-64.
- Reddy K. N. and Rebeiz, C. A. (1994). Modulators of the porphyrin pathway beyond protox. In: Porphyric Pesticides : Chemisry,
Toxicology, and Pharmaceutical Applications (S. O. Duke and C. A. Rebeiz, eds.) ACS Symposium Series 559, pp. 161-190.
- Gut, L. J., Juvik, J. A. and Rebeiz, C. A. (1994). Porphyric insecticides. In: Porphyric Pesticides: Chemisry, Toxicology, and
Pharmaceutical Applications (S. O. Duke and C. A. Rebeiz, eds.) ACS Symposium Series 559. pp. 206-232.
- Rebeiz, N., Kelley, K. W. and Rebeiz, C. A. (1994). Porphyrins as chemotherapeutic agents: Biochemistry of protoporphyrin IX
accumulation in mammalian cells. In: Porphyric Pesticides: Chemistry, Toxicology, and Pharmaceutical Applications (S. O. Duke and
C. A. Rebeiz, eds.) ACS Symposium Series 559. pp. 233-246.
- Rebeiz, C. A., Gut, L. J., Lee, K., Juvik, J. A., Rebeiz, C. C., Bouton, C. E. (1995) Photodynamics of porphyric insecticides. Crit.
Rev. Plant Sci. 14: 329-366.
- Parham R. and Rebeiz, C. A. (1995) Chloroplast Biogenesis 72: A [4-Vinyl]chlorophyllide a Reductase assay using divinyl chlorophyllide a
as an exogenous substrate. Anal Biochem. 231: 164-169.
- Lee K, and Rebeiz, C. A. (1995) Subcellular localization of protoporphyrin IX and its photodynamic effects on mitochondrial function of
the cabbage looper (Trichoplusia ni). In: Light Activated Pesticides (J. R. Heitz and K. R. Downum, eds.) ACS Symposium Series 616
, pp 152-163.
- Amindari S., Splittstosser, W. E., and Rebeiz C. A. (1995) Photodynamic effects of several metabolic tetrapyrroles on isolated
chloroplasts. In: Light Activated Pesticides (J. R. Heitz and K. R. Downum, eds.) ACS Symposium Series 616, pp 217-246.
- Kim, Jin-Seog, and Rebeiz, C. A. (1995). An improved analysis for determination of monovinyl and divinyl protoporphyrin IX.
J. Photosci. 2: 103-106.
- Rebeiz, N., Arkins, S. Rebeiz, C. A., Simon, J., Zachary, J. F., and Kelley, K. W. (1996). Induction of tumor necrosis by
delta-aminolevulinic acid and 1,10-phenanthroline. Cancer Res. 56:339-344.
- Fasoula, D. A., Smyth, C. and Rebeiz, C. A. (1996) Relationship of the Protochlorophyllide a content to Plant yield. In:
Handbook of Photosynthesis, (M. Pessarakli, ed.) pp 671-679, Marcel Dekker, New York.
- Rebeiz, N., Arkins, S., Kelley, K. W., and Rebeiz, C. A. (1996). Enhancement of coproporphyrinogen III transport into isolated
transfromed leukocyte mitochondria by ATP. Arch. Biochem. Biophys. 333:475-481.
- Kim, Jin-Seog, and Rebeiz, C. A. (1996) Origin of the chlorophyll a biosynthetic heterogeneity in higher plants. J. Biochem.
Mol. Biol. 29: 327-334.
- Awad Abd El Mageed, H., El Sahhar, K. F., Robertson, K. R., Parham, R. and Rebeiz, C. A. (1997) Chloroplast Biogenesis 77: Two novel
monovinyl and divinyl light-dark greening groups of plants and their relationships to the Chlorophyll a biosynthetic heterogeneity
of green plants. Photochem. Photobiol. 66:86-96.
- Ioannides I. M., Shedbalkar, V. P., and Rebeiz, C. A. (1997) Quantitative determination of 2-monovinyl protochlorphyll)ide) b
by spectrofluorometry. Anal. Biochem. 249:241-244.
- Kim, J. S., Kolossov, V. and Rebeiz, C. A. (1997). Chloroplast biogenesis 76. Regulation of 4-vinyl reduction during conversion of divinyl
Mg-protoporphyrin IX to monovinyl protochlorophyllide a is controlled by plastid and membrane stromal factors. Photosynthetica 34: 569-58.
-
Adra, A. N. and Rebeiz, C. A. (1998) Chloroplast Biogenesis 81. Tansient Formation of divinyl chlorophyll a following a 2.5 millisecond
light flash treatment of etiolated cucumber cotyledons. Photochem. Photobiol. 68: 852-856 .
- Rebeiz, C. A., Ioannides, I. M., Kolossov, V. and Kopetz, K. J. (1999) Chloroplast Biogenesis 80. Proposal of a unified chlorophyll
a/b biosynthetic pathway. Photosynthetica. 36:117-128 .
- Kolossov, V., Ioannides, I. M., Kulur, S., and Rebeiz, C. A.. (1999) Chloroplast Biogenesis 82. Development of a Cell-Free System
Capable of the Net Synthesis of Chlorophyll(ide) b. Photosynthetica. 36: 253-258 .
- Rebeiz, N., Arkins, S., Kelley, K. W., Dura, G., and Rebeiz, C. A.. (2001) . Modulator of heme biosynthesis induces apoptosis in
leukemia cells. J. Clin. Laser Surgery.. 19: 59-67 .
- Kolossov, V. L., and Rebeiz, C. A. (2001). Chloroplast Biogenesis 84: Solubilization and partial purification of membrane-bound [4-vinyl]
chlorophyllide a reductase from etiolated barley leaves Anal. Biochem.. 295: 214-219.
- Rebeiz, C. A. (2002). Analysis of Intermediates and end products of the chlorophyll biosynthetic pathway. In Heme, Chlorophyll, and
Bilins: Methods and Protocols, (A. G. Smith, and M. Witty eds.) Humana press , pp 111-155.
- Kolossov, V. Kopetz, K. J., and Rebeiz, C. A. (2003). Chloroplast Biogenesis 87: Evidence of resonance excitation energy transfer between
tetrapyrrole intermediates of the chlorophyll biosynthetic pathway and chlorophyll a. Phtochem. Photobiol. 78: 184-196.
- Rebeiz, C. A. Kolossov, V. L., Briskin, D., and Gawienowski, M. (2003). Chloroplast Biogenesis:
Chlorophyll biosynthetic heterogeneity, multiple biosynthetic routes and biological spin-offs. In, Handbook of Photochemistry and
Photobiology. (H. S. Nalwa ed.), American Scientific Publishers, Los Angeles, Vol 4, pp. 183-248.
- Kolossov V. L. and Rebeiz, C. A. (2003) Chloroplast Biogenesis 88: Protochlorophyllide b
Occurs in Green but not in Etiolated Plants. J. Biol. Chem. 278: 49675-49678
- Kopetz, K. K., Kolossov, V. L., and Rebeiz, C. A. (2004). Chloroplast Biogenesis 89: Development of analytical tools for probing the
biosynthetic topography of photosynthetic membranes by determination of resonance excitation energy transfer distances separating metabolic
tetrapyrrole donors from chlorophyll a acceptors. Anal. Biochem. 329:207-219 (Download a reprint)
- Rebeiz, C. A., Kolossov, V. L. and Kopetz, K. K. (2004). Chloroplast Bioengineering 1. Photosynthetic efficiency, Modulation of the photosynthetic unit size, and the agriculture of the future. In: Agricultural Applications of Green Chemistry (W. M. Nelson, ed.) ACS Symposium Series 887 pp 81-105.(Download a reprint).
Kolossov V. L. and Rebeiz, C. A. (2005). Chloroplast Biogenesis 91: Detection of d-aminolevulinic acid esterase
activity in higher plant and insect tissues. Pest.Biochem. Physiol. 83: 9-20.
d-Aminolevulinic acid (ALA) esterase(s) is an enzyme or a family of enzymes that regenerate ALA from ALA esters
by hydrolysis. These enzyme(s) are highly active in cancer cells. As a consequence ALA esters have been used to advantage in ALA-dependent
photoradiation therapy, since ALA esters translocate better to sites of metabolism in cancer cells and tissues than free ALA. In this work it
is shown that ALA esterase(s) also occur in insect and plant tissues, but are less active than in cancer cells. In plant cells ALA esterase
activity is observed in the cytosol as well as in the plastids where most of the activity is observed in the plastid stoma. The ALA esterase
activity appears to be sensitive to the nature of the esterifying alcohol as well as to components of the incubation medium. The observed
lower activity of ALA ester conversion to tetrapyrroles in insect and plant cells, in comparison to free ALA, suggests that the use of ALA
esters in photodynamic insecticidal and herbicidal applications may not be as advantageous as their use in cancer photodynamic therapy
treatments. It is proposed that ALA esterase(s) may be involved in the mobilization of sequestered and esterified ALA. Esterification and
sequestering of excess ALA may be visualized as a mean of cellular detoxification.
Kolossov V. L., Bohnert H., and Rebeiz, C. A. (2006) Chloroplast biogenesis 92: In situ screening for divinyl chlorophyll(ide) a
reductase mutants by spectrofluorometry. Anal. Biochem. In Press.
Chlorophyll biosynthetic heterogeneity is mainly rooted in parallel divinyl (DV) and monovinyl (MV) biosynthetic routes interconnected by
[4-vinyl] reductases (4VR), that convert DV tetrapyrroles to MV tetrapyrroles by conversion of the vinyl group at position 4 of the macrocycle
to ethyl. What is not clear at this stage is whether the various 4VR activities are catalyzed by one enzyme of broad specificity or by a
family of enzymes encoded by one or multiple genes, and each enzyme having narrow specificity. Additional research is needed to identify the
various regulatory components of 4-vinyl reduction. In this undertaking, Arabidopsis mutants that accumulate DV chlorophyllide and/or
DV chlorophyll [Chl(ide)] a are likely to provide an appropriate resource. Since the Arabidopsis genome has been completely sequenced,
the best strategy for identifying 4VR and/or putative regulatory 4VR genes is by screening Arabidopsis Chl mutants, for DV Chl(ide) a
accumulation. In wild type Arabidopsis, a DV plant species, only MV chlorophyllide (Chlide) a is detectable. However in Chl mutants lacking
4VR activity, DV Chl(ide) a may accumulate in addition to MV Chl(ide) a. In this work, an in situ assay of DV Chl(ide) a accumulation,
suitable for screening large number of mutants, lacking 4-vinyl Chlide a reductase activity with minimum experimental handling is described.
The assay involves homogenization of the tissues in Tris-HCl:Glycerol buffer and the recording of Soret excitation spectra at 77K. DV Chlide a
formation is detected by a Soret excitation shoulder at 459 nm over a wide range of DV Chlide a/MV Chl a ratios. The DV Chlide a shoulder
became undetectable below DV Chlide a/MV Chl a ratios of 0.049, i.e. below a DV Chlide a content of 5%.
Rebeiz, C. A. Kopetz, K. J. and Kolossov, V. L. (2006). Chloroplast Biiogenesis 90. Probing the Relationship between chlorophyll
biosynthetic routes and the topography of chloroplast biogenesis by resonance excitation energy transfer determinations. In: handbook pof
Photosynthesis(M. Pessarakli, ed.) Marcel Dekker Inc. New York. In press (Download a preprint)
The thorough understanding of photosynthetic membrane assembly requires a deeper knowledge of the coordination and regulation of the
chlorophyll (Chl) and thylakoid apoprotein biosynthetic pathways. As a working hypothesis we have recently proposed three different
chlorophyll (Chl)-thylakoid apoprotein biosynthesis models, namely: a single-branched Chl biosynthetic pathway (SBP)-single location model,
a SBP- multilocation model and a multibranched Chl biosynthetic pathway (MBP)-sublocation model [Handbook
of Photochemistry and Photobiology (2003), Am. Sc. Publishers, Los Angeles, pp 183-248]. The detection of resonance excitation energy
transfer between tetrapyrrole precursors of Chl, and several Chl-protein complexes [Photochem. Photobiol.
(2003) 78: 184-196] makes it possible to test the validity of the proposed Chl-thylakoid apoprotein biosynthesis models by resonance
excitation energy transfer determinations. In this work, the development of analytical tools that allow the detection of resonance excitation
energy transfer in green tissues, in situ, as well as the determination of resonance excitation energy transfer rates, and distances
separating tetrapyrrole donors from Chl-protein acceptors in green plants, by using readily available electronic spectroscopic instrumentations
are reviewed. It is concluded that the calculated distances are compatible with the MBP-sublocation model, and are incompatible with the
operation of the SBP-single location Chl-protein biosynthesis model.
