Nanobiotechnology Research at UTS

STELLA VALENZUELA HTTPS://WWW.UTS.EDU.AU/STAFF/STELLA.VALENZUELA

• Molecular and Cell Biologist working in the area of Bionanotechnology • Joined UTS 2001, previously worked at UNSW Faculty of Medicine,

Centre for Immunology, St Vincent’s Hospital, Cellabs P/L and Australian Monoclonal Development P/L

• Current industry partner - Surgical Diagnostics P/L together since 2004 held 3 ARC Linkage projects ($0.7M ARC funding) currently hold a joint ARC DP grant with SDX and UNSW and part of new ARC Industry Transformation Research Hub ($3.7M ARC funding) • New industry partner – BOD Australia P/L • Other collaborators BRAGG Institute ANSTO, UNSW, Macquarie Uni

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The scale of things: Nanoscale objects are any structures that are <100nm in size.

www.nano.gov

All biological systems operate via a process of molecular recognition; for example: hormones, cytokines, antibodies, neurotransmitters, ion channel gating, gene expression. Bionanotechnology Research at UTS has at its core the study of the molecular basis of biological recognition mechanisms:

• functional / kinetics studies • structural studies

Applications: • drug delivery • therapeutic applications • pesticides • implantable devices

Nanobiotechnology

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Nanomaterials Nanodevices

Nanocharacterisation

Nanobiotechnology

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“SMART” LIPOSOMAL SYSTEMS

Lipid coated

polyelectrolyte capsules

Non-lipid coated polyelectrolyte

capsules

Lipid coated polyelectrolyte

capsules

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RAW cells+gold+laser RAW cells+laser

TARGETED CELL KILLING USING GOLD CONJUGATED ANTIBODIES AND EXPOSURE TO PHOTOTHERMAL

ENERGY

Dakrong Pissuwan, Stella M. Valenzuela and Michael B. Cortie. (2006) Therapeutic possibilities of plasmonically heated gold nanoparticles. Trends in Biotechnology. 24(2): 62-67.

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TEM image of functionalised gold nanoparticles attached to

Toxoplasma gondii parasites

TARGETED CELL KILLING USING GOLD CONJUGATED ANTIBODIES AND EXPOSURE TO PHOTOTHERMAL

ENERGY

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science.uts.edu.au 11

(A) AFM error mode image of S100A8/A9 protein complex associated with lipid (the complex with the lipid had been preincubated at 37oC). The protein complex ranged from 35 to 45 nm in diameter; (B) Cross section of S100A8/A9 in lipid revealed a lipid height of 1.3 nm and a protein height of 5.3 nm; (C) AFM height mode image of S100A8/A9 protein complex preferentially attached to the edges of the lipid “patches”.

Stella M. Valenzuela, Mark Berkahn, Donald K. Martin, Thuan Huynh, Zheng Yang, Carolyn L. Geczy. (2006) Elucidating the Structure and Function of S100 Proteins in Membranes. Proc. of SPIE: BioMEMS and Nanotechnology II. Volume 6036: 603619-1.

LIPID-MEMBRANE TECHNOLOGIES…

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S100A9 Protein Imaged in Buffer Using Tapping Mode AFM

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CLIC1-CLIC6

Vertebrate CLICs

Invertebrate CLICs

EXC-4, EXL-1 DmCLIC

Littler et al FEBS 584:2093 (2010)

CLICs are highly conserved across vertebrates with related CLIC-like proteins in invertebrates

Extracellular fluid

Intracellular fluid

Plasma Membrane

CLIC1 proteins

Transmembrane form

Channel form

Factors that regulate the membrane insertion of CLIC1:

Redox environment Low pH

Membrane lipid composition

NOT A TYPICAL ION CHANNEL PROTEINS -

SPONTANEOUS MEMBRANE INSERTION OF SOLUBLE CLIC1

Harrop S et al, J Biol Chem , 276(48):44993-45000.

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Protein

PROTEIN CONDUCTANCE MEASUREMENTS USING TETHERED MEMBRANES AND IMPEDANCE

SPECTROSCOPY

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Conductance of CLIC1 in tBLMs containing varying ratios of cholesterol

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Langmuir monolayer films

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Effect of cholesterol on CLIC1 protein interactions with mixed lipid monolayers

Spontaneous Membrane Inserting Proteins

Bacterial toxins like listeriolysin, perfringolysin, streptolysin Pneumolysin

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ACKNOWLEDGEMENTS

Dr Heba Al Khamici Prof Mike Cortie Ms K Rufaka Hossain Ms Hala Ali Dr Lele Jiang A/Prof Mary Davey Dr Josh Chou Prof Don Martin Mr Mark Berkhan Prof Bruce Cornell Dr Stephen Holt BRAGG Institute ANSTO Prof Paul Curmi UNSW Dr Victoria Timchenko UNSW Dr Louise Brown Macquarie University Prof Sam Breit St Vincent’s Hospital Sydney