In Vitro Reconstitution and Biophysical Characterization of OEP16, an Outer Envelope Pore Protein of Pea Chloroplasts†

Biochemistry ◽  
2000 ◽  
Vol 39 (36) ◽  
pp. 11050-11056 ◽  
Author(s):  
Dirk Linke ◽  
Joachim Frank ◽  
Josef F. Holzwarth ◽  
Jürgen Soll ◽  
Christoph Boettcher ◽  
...  
Keyword(s):  
Outer Envelope ◽  
Biophysical Characterization ◽  
In Vitro Reconstitution ◽  
Pore Protein

scholarly journals Biochemical and Biophysical Characterization of a Chimeric TRIM21-TRIM5α Protein

2008 ◽  
Vol 82 (23) ◽  
pp. 11669-11681 ◽  
Author(s):  
Alak Kanti Kar ◽  
Felipe Diaz-Griffero ◽  
Yuan Li ◽  
Xing Li ◽  
Joseph Sodroski
Keyword(s):  
Variable Region ◽  
Globular Structure ◽  
Biophysical Characterization ◽  
Immunodeficiency Virus ◽  
Infected Insect ◽  
Low Levels ◽  
Trim Proteins

ABSTRACT The tripartite motif (TRIM) protein, TRIM5α, is an endogenous factor in primates that recognizes the capsids of certain retroviruses after virus entry into the host cell. TRIM5α promotes premature uncoating of the capsid, thus blocking virus infection. Low levels of expression and tendencies to aggregate have hindered the biochemical, biophysical, and structural characterization of TRIM proteins. Here, a chimeric TRIM5α protein (TRIM5Rh-21R) with a RING domain derived from TRIM21 was expressed in baculovirus-infected insect cells and purified. Although a fraction of the TRIM5Rh-21R protein formed large aggregates, soluble fractions of the protein formed oligomers (mainly dimers), exhibited a protease-resistant core, and contained a high percentage of helical secondary structure. Cross-linking followed by negative staining and electron microscopy suggested a globular structure. The purified TRIM5Rh-21R protein displayed E3-ligase activity in vitro and also self-ubiquitylated in the presence of ubiquitin-activating and -conjugating enzymes. The purified TRIM5Rh-21R protein specifically associated with human immunodeficiency virus type 1 capsid-like complexes; a deletion within the V1 variable region of the B30.2(SPRY) domain decreased capsid binding. Thus, the TRIM5Rh-21R restriction factor can directly recognize retroviral capsid-like complexes in the absence of other mammalian proteins.

Biophysical Characterization of SG2NA Variants and their Interaction with DJ-1 and Calmodulin in vitro

2018 ◽  
Vol 76 (4) ◽  
pp. 451-461 ◽  
Author(s):  
Sangeeta Soni ◽  
Buddhi Prakash Jain ◽  
Richa Gupta ◽  
Sudhaker Dharavath ◽  
Karunakar Kar ◽  
...  
Keyword(s):  
Biophysical Characterization

In vitro Reconstitution and Characterization of the Yeast Mitochondrial Degradosome Complex Unravels Tight Functional Interdependence

2007 ◽  
Vol 372 (1) ◽  
pp. 23-36 ◽  
Author(s):  
Michal Malecki ◽  
Robert Jedrzejczak ◽  
Piotr P. Stepien ◽  
Pawel Golik
Keyword(s):  
In Vitro Reconstitution
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