Abstract

Ile caseinolytic protease from E. coli is comprised of two subunits ihe protease, CIpP, and the ATPase, CIpA. Fully active CIpP has been over-expressed in R coli and purified to homogeneity. Mass measurements from the STEM indicate that it is a tetradecamer. In stain, it appears as two stacked seven-member rings. Small angle X-ray and neutron scattering curves were determined for ClpP in solution. A model was derived from these data of a cylinder with an axial pore. Ile size of the pore is similar to that found in chaperone proteins. CIpP has been crystallized and the above data. has been used to help solve the x-ray structure. Several cysteine mutants were engineered and Au-labeled and examined in the STEK to aid in the determination of the structure. CIpA has been examined in the STEM. Preliminary samples were not very good but finther attempts with better purified samples are underway. The CIpA component may be hexameric. The final goal is to look at the interaction of CIpP and CIpA.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001777-19
Application #
6444693
Study Section
Project Start
2001-04-01
Project End
2002-09-29
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
19
Fiscal Year
2001
Total Cost
$293,069
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; 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
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
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
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

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