Laser Ablation Capture Mass Spectrometry for Single Cell Proteomics

LSU Chemistry Professor Secures NIH Grant for Single Cell Analysis Research

LSU Research News

Grant is one of 15 high risk, high impact projects funded nationally

Kermit Murray, LSU professor of chemistry, was awarded $350,844 to develop new methods for single cell analysis. The goal of Murray’s project, entitled, Nanoscale Laser Ablation Capture Mass Spectrometry for Single Cell Proteomics, is to create nanometer scale sampling for mass spectrometry using near-field optics and laser ablation.

“An ensemble of cells does not give us a detailed look at cells,” said Murray. “You may have 100 cells and only a few of them will be in a particular disease state. The use of mass spectrometry will give us a high-resolution view of cells at a sub-cellular level.”

Single cell analysis emerged as an important field of research after new technologies with improved sensitivity made it possible to measure cell-to-cell differences in living organisms and correlate the variation with changes in biological function and disease.

Murray’s research was one of only 15 high-risk/high-impact projects funded nationally to develop new tools to enhance measurement parameters such as sensitivity, selectivity, spatiotemporal resolution, scalability and/or non-destructive measures that preserve the integrity of the cell. The new tools will also improve capabilities for the simultaneous measurement of multiple molecular components, such as genes and cells, within a single cell.

Advancements in single cell analysis will allow personalized health care at the cellular level. Research could lead to the development of techniques to identify genomic lesions in individual cancer cells, which would improve the early detection of rare tumor cells, allow better monitoring of circulating tumor cells and guide chemotherapy treatments.

“This was an extremely competitive grant program. I was really surprised that we scored as well as we did,” said Murray. “It is a very prestigious award and I am glad to bring such quality research to LSU and the state of Louisiana.”

The Single Cell Analysis Program is funded through the Common Fund, and managed by the NIH Office of the Director in partnership with the National Institute of Biomedical Imaging and Biotechnology, or NIBIB, and National Institute of Mental Health, or NIMH, both part of NIH.

The NIH Common Fund encourages collaboration and supports a series of exceptionally high impact, trans-NIH programs. Common Fund programs are designed to pursue major opportunities and gaps in biomedical research that no single NIH Institute could tackle alone, but that the agency as a whole can address to make the biggest impact possible on the progress of medical research. Additional information about the NIH Common Fund can be found at http://commonfund.nih.gov.

NIBIB’s mission is to support multidisciplinary research and research training at the crossroads of engineering and the biological and physical sciences. NIBIB supports emerging technology research and development within its internal laboratories and through grants, collaborations, and training. More information is available at the NIBIB website: http://www.nibib.nih.gov/.

The mission of NIMH is to transform the understanding and treatment of mental illnesses through basic and clinical research, paving the way for prevention, recovery and cure. More information is available at the NIMH Web site, http://www.nimh.nih.gov.

About the National Institutes of Health:

NIH, the nation’s medical research agency, includes 27 Institutes and Centers and is a component of the U.S. Department of Health and Human Services. NIH is the primary federal agency conducting and supporting basic, clinical, and translational medical research, and is investigating the causes, treatments, and cures for both common and rare diseases. For more information about NIH and its programs, visit http://www.nih.gov/.

Choppin Annex Interior

Choppin Annex front hall looking out onto Highland Road
Front hall looking out onto Highland Road
Choppin Annex fume hoods.
Fume hoods in a first floor instrument lab
Choppin Annex conference room
All floors have a conference room (except floor 5 which is currently unfinished)
Choppin Annex 5th floor
The 5th floor of the Choppin Annex is currently a shell and will be outfitted for materials analysis in the near future.

ASMS 2012

Murray Group at the poster session at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Venkatesulu Salla at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Machavaram Siva Pratap Reddy at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Chinthaka Seneviratne and Sung-Gun Park at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Yonathan Merid at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Murray Group at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Float Plane in Vancouver Harbor at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Float plane and boat at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Float planes at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Float plane taking off in Vancouver Harbor at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Two float planes in Vancouver Harbor at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012
Rainbow at the American Society for Mass Spectrometry meeting in Vancouver BC, 2012

Chapter 3 Links

Here are some examples of videos related to the content of Chapter 3 of Chemistry: the Central Science. You may find some of them a useful supplement to the assigned class material.

Stoichiometry

[youtube=http://www.youtube.com/watch?v=OjnuNRivedM]

Balancing Equations

[youtube=http://www.youtube.com/watch?v=YAutl-G7g4A] [youtube=http://www.youtube.com/watch?v=RnGu3xO2h74]

The Mole

[youtube=http://www.youtube.com/watch?v=tEn0N4R2dqA] [youtube=http://www.youtube.com/watch?v=BTRm8PwcZ3U] [youtube=http://www.youtube.com/watch?v=AsqEkF7hcII]

Mass Composition to Empirical Formula

[youtube=http://www.youtube.com/watch?v=xatVrAh2U0E] [youtube=http://www.youtube.com/watch?v=sXOIIEZh6qg]

Combustion Analysis

[youtube=http://www.youtube.com/watch?v=ebFP7onBbts] [youtube=http://www.youtube.com/watch?v=PCtR5hqBWjM] [youtube=http://www.youtube.com/watch?v=66ziUq6vRko]

Gram to Mole to Mole to Gram Problems

[youtube=http://www.youtube.com/watch?v=jFv6k2OV7IU] [youtube=http://www.youtube.com/watch?v=eQf_EAYGo-k]

Limiting Reactants

[youtube=http://www.youtube.com/watch?v=krioEzlRegc] [youtube=http://www.youtube.com/watch?v=VBuCEwTwV7Q] [youtube=http://www.youtube.com/watch?v=GY2Mav0_mGU] [youtube=http://www.youtube.com/watch?v=rESzyhPOJ7I] [youtube=http://www.youtube.com/watch?v=CMi9k8ts9r4] [youtube=http://www.youtube.com/watch?v=YNriRslOk9A]

Isolation and determination of the primary structure of a lectin protein from the serum of the american alligator (Alligator Mississippiensis)

doi:10.1016/j.cbpb.2011.11.001
Darville, Merchant, Maccha, Siddavarapu, Hasan, and Murray
Comp. Biochem Physiol. B

Alligator Lectin Protein Sequence (doi:10.1016/j.cbpb.2011.11.001)

Mass spectrometry in conjunction with de novo sequencing was used to determine the amino acid sequence of a 35 kDa lectin protein isolated from the serum of the American alligator that exhibits binding to mannose. The protein N-terminal sequence was determined using Edman degradation and enzymatic digestion with different proteases was used to generate peptide fragments for analysis by liquid chromatography tandem mass spectrometry (LC MS/MS). Separate analysis of the protein digests with multiple enzymes enhanced the protein sequence coverage. De novo sequencing was accomplished using MASCOT Distiller and PEAKS software and the sequences were searched against the NCBI database using MASCOT and BLAST to identify homologous peptides. MS analysis of the intact protein indicated that it is present primarily as monomer and dimer in vitro. The isolated 35 kDa protein was ~ 98% sequenced and found to have 313 amino acids and nine cysteine residues and was identified as an alligator lectin. The alligator lectin sequence was aligned with other lectin sequences using DIALIGN and ClustalW software and was found to exhibit 58% and 59% similarity to both human and mouse intelectin-1. The alligator lectin exhibited strong binding affinities toward mannan and mannose as compared to other tested carbohydrates.

Experimental Physical Chemistry Tenure Track Position at LSU

ASSISTANT/ASSOCIATE/FULL PROFESSOR

The Louisiana State University Department of Chemistry anticipates filling an Assistant/Associate/Full Professor (Experimental Physical Chemistry/Tenure-track) position with a starting date of August 13, 2012.

Required Qualifications: Ph.D. in chemistry or a related field; demonstrated excellence in teaching and research. Responsibilities: establish a strong, well-funded, widely recognized research program in experimental physical chemistry; teach at the undergraduate and graduate levels. An offer of employment is contingent on a satisfactory pre-employment background check.

Application deadline is November 28, 2011 or until a candidate is selected.

Applications at the Assistant Professor level should consist of a cover letter, curriculum vitae, summary of proposed research, and statement of teaching philosophy, preferably as a single PDF document; and three letters of recommendation.

Associate/Full Professor candidates should submit a letter of interest and curriculum vitae. Please submit materials electronically; see below. Arrange for letters of recommendation to be sent to Ms. Vickie Thornton (vthornton@lsu.edu), Experimental Physical Chemistry Search, Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803.

LSU IS AN EQUAL OPPORTUNITY/EQUAL ACCESS EMPLOYER

Apply online at https://lsusystemcareers.lsu.edu/applicants/Central?quickFind=53755