Talaga Group
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and protein conformational dynamics
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Talaga Group research to be featured on JMB cover.
The Journal of Molecular Biology has selected Jason Giurleo's paper on amyloid formation from ß-lactoglobulin for the cover of their September 19, 2008 issue. Congratulations Jason!
Link to published version
Jason T. Giurleo, Xianglan He and David S. Talaga, "β-Lactoglobulin Assembles into Amyloid through Sequential Aggregated Intermediates" J. Mol. Biol. (2008) 381:5
We have investigated the aggregation and amyloid fibril formation of bovine β-lactoglobulin variant A, with a focus on the early stages of aggregation. We used noncovalent labeling with thioflavin T and 1-anilino-8-naphthalenesulfonate to follow the conformational changes occurring in β-lactoglobulin during aggregation using time resolved luminescence. 1-Anilino-8-naphthalenesulfonate monitored the involvement of the hydrophobic core/calyx of β-lactoglobulin in the aggregation process. Thioflavin T luminescence monitored the formation of amyloid. The luminescence lifetime distributions of both probes showed changes that could be attributed to conformational changes occurring during and following aggregation. To correlate the luminescence measurements with the degree of aggregation and the morphology of the aggregates, we also measured dynamic light scattering and atomic force microscopy images. We evaluated the relative stability of the intermediates with an assay that is sensitive to aggregation reversibility. Our results suggest that initial aggregation during the first 5 days occurred with partial disruption of the characteristic calyx in β-lactoglobulin. As the globular aggregates grew from days 5 to 16, the calyx was completely disrupted and the globular aggregates became more stable. After this second phase of aggregation, conversion into a fibrillar form occurred, marking the growth phase, and still more changes in the luminescence signals were observed. Based on these observations, we propose a three-step process by which monomer is converted first into weakly associated aggregates, which rearrange into stable aggregates, which eventually convert into protofibrils that elongate in the growth phase.
Prof. Talaga Chairs session at Biophysical Society Meeting
Professor Talaga chaired a platform session at the Biophysical Society meeting in Long Beach on the Aggregation Behavior of Proteins. Many interesting talks were presented.
Troy Messina takes faculty position
After a busy and successful academic job search, Dr. Messina has decided to take a position as an Assistant Professor at Centenary College of Louisiana. We all wish him the best of luck and will miss him in the Talaga Labs.
NIRT: Ligand Nanodisplay for Cellular Internalization and Super-Activation
The NSF has awarded a 4-year grant to a Rutgers Nanoscale Interdisciplinary Research Team, or NIRT to study "Ligand Nanodisplay for Cellular Internalization and Super-Activation."
This NIRT proposal has assembled an interdisciplinary team of five principal/co-principal investigators from Rutgers and Princeton; and two supportive collaborators; to elucidate the significance of cellular internalization of nanodisplayed adhesion ligands toward cellular activation and motility. The proposed studies are predicated on a recent discovery by the PI that when traditionally non-internalizable adhesion ligands are displayed from truly nanoscale carriers (<100 nm), the adhesion ligands get endocytosed (internalized), and in the process of this cytointernalization, the ligands trigger greatly increased levels of intracellular biochemical signaling pathways, which leads to superactivation of cell behaviors, including cellular motility that is a major cellular phenomenon for biotechnology and tissue engineering. Because these substrate-based ligand nanocarriers are cleared and internalized by cells upon binding, the interfaces are biodynamic in nature.
The key questions addressed by the NIRT proposal are:
- How can such nanoscale displays be rationally engineered?
- What are their effects on intracellular molecular biosignaling?
- How can new ways be envisioned for biosensing of ligand-nanocarrier cell internalization?
Intellectual Merit: The proposed studies are innovative because they seek to engineer the presentation of well-established protein ligands via nanoscale carriers, in order to superactivate cells in specific ways; and to establish novel biosensing approaches for measurement of nanocarrier cytointernalization. Preliminary studies show that cells triggered to migrate using nanoscale cytointernalizable ligands were significantly more active than the controls, suggesting that nanotechnology can potentially help reveal new signaling domains or expanded signaling capabilities for our current repertoire of biomolecules. The design of a successful nanoscale biointerfacial system will involve the contributions from very diverse scientific professionals, including a cell/matrix biologist (Schwarzbauer) to engineer ligand fragments and examine their intracellular protein-level signaling; a cellular bioengineer (Moghe) to fabricate nanocarriers for the ligand display and quantify cellular function, uptake, and signaling; a molecular bioengineer (Roth) to screen intracellular signaling via parallel DNA microarrays; a physical spectroscopist (Talaga) to evaluate the intracellular trafficking; and inorganic nanomaterials scientist (Tsakalakos) and nanobiosensor expert (Menon) for developing sensors of nanocarrier uptake.
Broader Impact: Insights from the study can potentially impact the development of bioactive nanomaterials for cellular biotechnology; wound healing; tissue regeneration; and molecular biomedicine. The nanoscale platform identified herein can boost cell motility kinetics by an order-of-magnitude, which can reduce the time for wound healing and re-epithelialization, leading to reduced healthcare costs and improved therapies. At Rutgers, the NIRT proposal is particularly timely because it can further foster the growth of cross-disciplinary education and minority outreach in nanotechnology and life-sciences at graduate and undergraduate levels. Using existing partnerships with an existing IGERT program on biointerfaces directed by the PI, the Northeast Alliance for Graduate Education and the Professoriate (NEAGEP), and a tissue engineering postdoctoral training program, the NIRT team will launch a vigorous outreach effort to broaden participation at all levels (graduate, undergraduate, postdoctoral). Through the Rutgers NSF IGERT on biointerfaces (www.igert.rutgers.edu) (one major thrust is micro/nanoscale biointerfaces), the research themes of the NIRT will be disseminated among the undergraduate summer research group of minority students within the RISE (Research in Science and Engineering) program. The NIRT faculty are in various stages of developing three new research-integrative graduate courses, based on the NIRT theme, on cellular/molecular bioengineering; nano- and microsystems biointerfaces; and biointerfacial characterization. These courses will help establish a stronger educational infrastructure for nanoscale science and engineering, which synergizes with three advanced research centers (Keck Collaborative Center; Institute of Advanced Materials; NJ Center for Biomaterials) within New Jersey. The NIRT collaborations will facilitate the offering of an international internship to a NIRT graduate researcher at the Department of Micro/Nanosystems, Technical University, Denmark.
The Nanoscale Interdisciplinary Research Team:
Principal Investigator:
- P. Moghe (Cellular Bioengineering; Bio-Nanotechnology)
Co-PI's:
- C. Roth (Molecular Bioengineering)
- J. Schwarzbauer (Matrix Biology)
- D. Talaga (Molecular Imaging & Spectroscopy)
- T. Tsakalakos (Nanostructured Materials)
Collaborators:
- A. Menon (NanoBiosensors)
- J. Kohn (Polymeric Biomaterials)
Jeremy Pronchik awarded GAANN fellowship
Jeremy Pronchik awarded a GAANN fellowship as part of the NSF Computer Science, Engineering, and Mathematics Scholarships (CSEMS)
NSF Computer Science, Engineering, and Mathematics Scholarships (CSEMS) This program supports scholarships for academically talented, financially needy students, enabling them to enter the high technology workforce following completion of an associate, baccalaureate, or graduate level degree in computer science, computer technology, engineering, engineering technology, or mathematics. Academic institutions apply for awards to support scholarship activities, and are responsible for selecting scholarship recipients, reporting demographic information about student scholars, and managing the CSEMS project at the institution.
Huang Cheng-Yen joins the Talaga group
Dr. Huang is currently working on examining how hidden Markov models may be used to reconstruct Langevin dynamics.
Dr. Huang joins us after two years as a post-doc with Tom Spiro in the Princeton Chemistry Department. At Princeton Cheng-Yen was developing temperature-jump deep-UV resonance Raman techniques to measure protein folding and conformational dynamics
Cheng-Yen completed his Ph.D. in the group of Feng Gai at the University of Pennsylvania where he worked on uncovering the fast events in protein and peptide folding using temperature jump IR spectroscopy.