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Nicholas C Carpita

Botany and Plant Pathology 

  • Professor
765.494.4653
765.494.0363
Lilly Hall Room 1-464

Cellulose structure and biosynthesis

With more than 200 billion tons per year in the natural environment, cellulose is the most abundant biopolymer on Earth. Cellulose microfibrils, the fundamental scaffolding of the plant cell wall, is a para-crystalline array of several dozen (1->4)-b-D-glucan chains synthesized at the plasma membrane surface by large multicomponent complexes of cellulose synthase (CesA) proteins. We discovered that recombinant catalytic domains of CesA are two-domain structures that dimerize using Small-Angle X-ray Scattering (SAXS) experiments to derive 3-D surface contour structures (Olek et al. 2014). The catalytic domains of plant CesAs contain two unique sequences not found in prokaryotic ancestors ­– a Plant-Conserved Region (P-CR) and Class-Specific Region (CSR) of unknown function. Molecular docking experiments with the catalytic core predicted that the CSRs of CesAs are the dimerization domains. We aim to provide the first crystal structure of a plant CesA catalytic domain. Towards that goal, we crystallized a recombinant Plant-Conserved Region (P-CR) and showed that it is primarily a coiled-coil domain positioned near the entrance to the active site of the catalytic core (Rushton et al. 2016). With Wen Jiang (Purdue) we have begun studies to define the assembly of CesAs into complexes at the Golgi membrane as part of a broader effort to characterize the dynamics of the Golgi proteome.  From substrate binding stoichiometry, we know that each CesA protein synthesizes a single (1->4)-b-D-glucan chain of a microfibril (Olek et al. 2014). We developed a modified TEMPO-catalyzed oxidation of glucose to uronosyl residues followed by carboxyl reduced with NaBD4 to provide a novel method for determining proportion of disordered surfaced chains, ordered surface chains in which only one-half of the residues are exposed to water, and completely anhydrous core-crystalline residues for cellulose microfibrils in biomass. Microfibrils of primary or secondary walls from seedlings and lignocellulosic biomass have significantly higher content of their glucan chains in anhydrous domains, indicating cellulose microfibrils into bundles with extensive crystalline continuity.

 

Click here for Dr. Carpita's complete research summary - https://ag.purdue.edu/btny/Pages/Nick-Carpita-Research-summary.aspx

 

Awards & Honors

(2014) 2014 Team Award. College of Agriculture.

(2010) President of American Society of Plant Biologist. American Society of Plant Biologist.

(2015) Seeds of Success Award, Millionaire's Club. College of Agriculture.

(2009) Fellow of American Society of Plant Biologists. American Society of Plant Biologists.

(1991) Agricultural Research Award. Purdue University.

(1987) Gamma Sigma Delta. Purdue University.

(1984) Sigma Xi. Purdue University.

Patents

Carpita, N. C., Hodges, T. K., & Antunes, M. Benzoate inducible promoters and promoter systems are disclosed, and uses thereof. Polynucleotides disclosing Benzoate Response Elements are also disclosed.. U.S. Patent No. 07705203. Washington, D.C.: U.S. Patent and Trademark Office.

Selected Publications

Rushton, P. S., Olek, A. T., Makowski, L., Badger, J., Steussy, C. N., Carpita, N. C., & Stauffacher, C. V. (2017). Rice Cellulose SynthaseA8 Plant-Conserved Region Is a Coiled-Coil at the Catalytic Core Entrance. Plant Physiology, 173(1), 482-494. doi:10.1104/pp.16.00739

Dugard, C. K., Mertz, R. A., Rayon, C., Mercadante, D., Hart, C., Benatti, M. R., . . . Carpita, N. C. (2016). The Cell Wall Arabinose-Deficient Arabidopsis thaliana Mutant murus5 Encodes a Defective Allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2. Plant Physiology, 171(3), 1905-20. doi:10.1104/pp.15.01922

McCann, M. C., & Carpita, N. C. (2015). Biomass recalcitrance: A multi-scale, multi-factor and conversion-specific property. The Journal of Experimental Botany, 66, 4109-4118.

Penning, B., Sykes, R., Babcock, N., Dugard, C., Held, M., Klimek, J., . . . Carpita, N. (2014). Genetic determinants for enzymatic digestion of lignocellulosic biomass are independent of those for lignin abundance in a maize recombinant inbred population. Plant Physiology, 165, 1475-1487.

Olek, A. T., Rayon, C. J., Makowski, L., Kim, H., Ciesielski, P., Badger, J., . . . Carpita, N. C. (2013). ) Small-angle x-ray scattering reveals the structure of the catalytic domain of a cellulose synthase and its assembly into dimers. Plant Cell, 26, 2966-3009. doi:10.1105/tpc.114.126862

Rayon, C., Olek, A. T., & Carpita, N. C. (2013). Towards redesigning cellulose biosynthesis for improved bioenergy feedstocks. In Plants and Bioenergy (183-193). New York, NY: Springer. doi:10.1007/978-1-4614-9329-7_11

Held, M. A., Penning, B., Brandt, A. S., Kessans, S. A., Yong, W., Scofield, S. R., & Carpita, N. C. (2008). Small-interfering RNAs from natural antisense transcripts derived from a cellulose synthase gene modulate cell wall biosynthesis in barley. Proceedings of the National Academy of Sciences of the United States of America, 105(51), 20534-9. doi:10.1073/pnas.0809408105

Naran, R., Chen, G., & Carpita, N. C. (2008). Novel rhamnogalacturonan I and arabinoxylan polysaccharides of flax seed mucilage. Plant Physiology, 148(1), 132-41. doi:10.1104/pp.108.123513

Urbanowicz, B. (2004). Topology of the maize mixed-linkage (1→3),(1→4)-beta-D-glucan synthase at the Golgi membrane. Plant Physiology, 134, 758-768. Retrieved from http://www.ag.purdue.edu/btny/Documents/PlantPhysiology134-2.pdf

Vergara, C. E., & Carpita, N. C. (2001). Beta-D-glycan synthases and the CesA gene family: lessons to be learned from the mixed-linkage (1-->3),(1-->4)beta-D-glucan synthase. Plant Molecular Biology, 47(1-2), 145-60.

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