A Serendipitous Discover that in situ Proteolysis is Essential for the Crystallization of Yeast CPSF-100 (Ydh1p)

Journal Article · Sun Jan 01 04:00:00 EST 2006 · Acta Crystallographica Section F: Structural Biology and Crystallization Communications

DOI:https://doi.org/10.1107/S1744309106038152· OSTI ID:930089

Mandel, C; Gebauer, D; Zhang, H; Tong, L

The cleavage and polyadenylation specificity factor (CPSF) complex is required for the cleavage and polyadenylation of the 3'-end of messenger RNA precursors in eukaryotes. During structural studies of the 100 kDa subunit (CPSF-100, Ydh1p) of the yeast CPSF complex, it was serendipitously discovered that a solution that is infected by a fungus (subsequently identified as Penicillium) is crucial for the crystallization of this protein. Further analyses suggest that the protein has undergone partial proteolysis during crystallization, resulting in the deletion of an internal segment of about 200 highly charged and hydrophilic residues, very likely catalyzed by a protease secreted by the fungus. With the removal of this segment, yeast CPSF-100 (Ydh1p) has greatly reduced solubility and can be crystallized in the presence of a minute amount of precipitant.

• Research Organization: Brookhaven National Laboratory (BNL) National Synchrotron Light Source
• Sponsoring Organization: Doe - Office Of Science
• DOE Contract Number: AC02-98CH10886
• OSTI ID: 930089
• Report Number(s): BNL--80725-2008-JA
• Journal Information: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Journal Name: Acta Crystallographica Section F: Structural Biology and Crystallization Communications Vol. 62
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CLEAVAGE
CRYSTALLIZATION
MESSENGER-RNA
PENICILLIUM
PROTEINS
PROTEOLYSIS
REMOVAL
RESIDUES
SOLUBILITY
SOLUTIONS
SPECIFICITY
YEASTS
national synchrotron light source