Abstract

Cytoskeletal actin is organized into bundles and networks by actin crosslinking proteins. T-fimbrin (70 kDa) is the smallest member of a family of these proteins and binds actin filaments as a monomer. A 40 kDa fragment of T-fimbrin (called N375) contains two actin binding domains, two cysteines available for labeling (one significantly more reactive than the other), and an x-ray structure for much of it. Their cryoEM map shows F-actin decorated with N375 to 28A resolution. A Nanogold tag on N375 would determine its orientation and help align the x-ray and EM data. The N375 was labeled with Nanogold and examined in the STEM. The labeling seemed quite good, though the protein is too small to appear interesting as a freeze-dried specimen. Hopeflilly, it will be helpftil for their cryoEM analyses.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
2P41RR001777-15
Application #
2805045
Study Section
Project Start
Project End
Budget Start
1996-10-01
Budget End
1997-09-30
Support Year
15
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
Brookhaven National Laboratory
Department
Type
DUNS #
027579460
City
Upton
State
NY
Country
United States
Zip Code
11973

  Publications

Hind, Geoffrey; Wall, Joseph S; Várkonyi, Zsuzsanna et al. (2014) Membrane crystals of plant light-harvesting complex II disassemble reversibly in light. Plant Cell Physiol 55:1296-303
Garofalo, Robert; Mustanski, Brian; Johnson, Amy et al. (2010) Exploring factors that underlie racial/ethnic disparities in HIV risk among young men who have sex with men. J Urban Health 87:318-323
Joshi, V N; Mitra, D; England, M D et al. (2010) Large Covalently Linked Fluorescent and Gold Nanoparticle Immunoprobes. Microsc Microanal 16:966-967
Ackerson, Christopher J; Powell, Richard D; Hainfeld, James F (2010) Site-specific biomolecule labeling with gold clusters. Methods Enzymol 481:195-230
Hu, Minghui; Qian, Luping; Brinas, Raymond P et al. (2008) Gold nanoparticle-protein arrays improve resolution for cryo-electron microscopy. J Struct Biol 161:83-91
Garofalo, Robert; Mustanski, Brian; Donenberg, Geri (2008) Parents know and parents matter;is it time to develop family-based HIV prevention programs for young men who have sex with men? J Adolesc Health 43:201-4
Brinas, Raymond P; Hu, Minghui; Qian, Luping et al. (2008) Gold nanoparticle size controlled by polymeric Au(I) thiolate precursor size. J Am Chem Soc 130:975-82
Hu, Minghui; Zhang, Yian-Biao; Qian, Luping et al. (2008) Three-dimensional structure of human chromatin accessibility complex hCHRAC by electron microscopy. J Struct Biol 164:263-9
Mustanski, Brian; Garofalo, Robert; Herrick, Amy et al. (2007) Psychosocial health problems increase risk for HIV among urban young men who have sex with men: preliminary evidence of a syndemic in need of attention. Ann Behav Med 34:37-45
Garofalo, Robert; Mustanski, Brian S; McKirnan, David J et al. (2007) Methamphetamine and young men who have sex with men: understanding patterns and correlates of use and the association with HIV-related sexual risk. Arch Pediatr Adolesc Med 161:591-6

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