This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. It is well recognized that the Fourier amplitudes of cryo-EM data are not as accurate as the corresponding phases. In general, low resolution amplitudes tend to be overweighted and high resolution amplitudes tend to be underweighted. The early work of Crowther showed that sharpening a map using a Gaussian function made the helices more pronounced in their 7.4 ? map of the hepatitis B capsid core (B?ttcher et al., 1997). A similar approach has been adopted in many of the reconstruction maps (e.g. (Zhang et al., 2008b)). However, the actual B factor parameter used for sharpening those maps was somewhat ad hoc. In the EMAN procedure, when we merge the data in 2-D to make class-averages, we apply a Wiener filter (Ludtke et al., 2004). When 3-D reconstruction of these class-averages is performed, this Wiener filter is no longer fully accurate due to the additional averaging that occurs during reconstruction. This results in a moderate overfiltration of EMAN reconstructions before post-processing, even if all of the CTF parameters have been determined accurately. Similarly, the MPSA algorithm also includes a weighting factor of the experimental B factor for each particle image when they are merged through a modified EMAN 3D reconstruction program (Liu et al., 2007b). To compensate, we generally perform a refiltering (sharpening) of the map prior to final interpretation and publication. We are assessing different approaches as how to sharpen the map to bring out the hidden features.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR002250-24
Application #
7953781
Study Section
Special Emphasis Panel (ZRG1-BPC-K (40))
Project Start
2008-12-01
Project End
2009-11-30
Budget Start
2008-12-01
Budget End
2009-11-30
Support Year
24
Fiscal Year
2009
Total Cost
$34,780
Indirect Cost
Name
Baylor College of Medicine
Department
Physiology
Type
Schools of Medicine
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030
Bucero, Marta Abril; Bajaj, Chandrajit; Mourrain, Bernard (2016) On the construction of general cubature formula by flat extensions. Linear Algebra Appl 502:104-125
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Baker, Mariah R; Fan, Guizhen; Serysheva, Irina I (2015) Single-particle cryo-EM of the ryanodine receptor channel in an aqueous environment. Eur J Transl Myol 25:35-48

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