Event Title

Purification and crystallization of I-SSC1

Research Mentor(s)

Paul Spiegel

Description

Homing endonucleases are a class of enzymes capable of hydrolyzing DNA within the nucleus of the cell that synthesized them. As the DNA is repaired the sequence encoding the endonuclease is often copied to the sight of cleavage. These target DNA sequences are commonly 12 to 40 base pairs long and differentiate the endonucleases into families. The 300 amino acid protein, I-SSC1 is an intron-encoded protein within the LAGLIDADG family, named from their target sequences. To date there are no proposed structures for this protein. We have been working on the purification and crystallization of a His-tagged version of I-SSC1. Expression and purification methods have been successfully determined using E. coli as a means of expression, and immobilized metal ion affinity chromatography (IMAC) for the purification. Crystallization conditions are still being optimized, but initial crystals have been grown.

Document Type

Event

Start Date

May 2018

End Date

May 2018

Location

Chemistry

Rights

Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this document for commercial purposes, or for financial gain, shall not be allowed without the author’s written permission.

Language

English

Format

application/pdf

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May 17th, 9:00 AM May 17th, 12:00 PM

Purification and crystallization of I-SSC1

Chemistry

Homing endonucleases are a class of enzymes capable of hydrolyzing DNA within the nucleus of the cell that synthesized them. As the DNA is repaired the sequence encoding the endonuclease is often copied to the sight of cleavage. These target DNA sequences are commonly 12 to 40 base pairs long and differentiate the endonucleases into families. The 300 amino acid protein, I-SSC1 is an intron-encoded protein within the LAGLIDADG family, named from their target sequences. To date there are no proposed structures for this protein. We have been working on the purification and crystallization of a His-tagged version of I-SSC1. Expression and purification methods have been successfully determined using E. coli as a means of expression, and immobilized metal ion affinity chromatography (IMAC) for the purification. Crystallization conditions are still being optimized, but initial crystals have been grown.