Presentation Title

Site- directed mutagenesis of key breast cancer residues in lysyl oxidase

Presenter Information

Carmen MezaFollow

Faculty Mentor

Karlo Lopez

Start Date

23-11-2019 8:00 AM

End Date

23-11-2019 8:45 AM

Location

31

Session

poster 1

Type of Presentation

Poster

Subject Area

behavioral_social_sciences

Abstract

Lysyl oxidase is a copper-dependent amine oxidase that has a key role in the cross-linkage of collagen and elastin. In addition, it has been shown that lysyl oxidase in its mature form is involved in metastasis. In this study a Nus-A tag was used to overcome the enzyme’s low solubility. Seven mutations will be introduced into the mature form of the enzyme. These mutations are linked to lysyl oxidase found in cancer cells and were selected because the mutated residues are conserved across all lysyl oxidase isoforms. The purpose of this study is to elucidate the role of these mutations in the enzyme and also to incorporate unique findings into breast cancer studies. At this point two mutations have been generated pQ293R and pC412S. However, pC412S is still in the process of being overexpressed and purified in order to be tested. The total enzymatic yield for pQ293R was 13.124 mg (6.607 mg/L of media) and had an enzymatic activity of 0.061 U/mg. This value show that the mutated enzyme has less activity than the wildtype, which has an activity value of 0.11 U/mg. Additional work is underway to determine if these mutations affect the ability of the enzyme to incorporate copper or to generate LTQ.

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Nov 23rd, 8:00 AM Nov 23rd, 8:45 AM

Site- directed mutagenesis of key breast cancer residues in lysyl oxidase

31

Lysyl oxidase is a copper-dependent amine oxidase that has a key role in the cross-linkage of collagen and elastin. In addition, it has been shown that lysyl oxidase in its mature form is involved in metastasis. In this study a Nus-A tag was used to overcome the enzyme’s low solubility. Seven mutations will be introduced into the mature form of the enzyme. These mutations are linked to lysyl oxidase found in cancer cells and were selected because the mutated residues are conserved across all lysyl oxidase isoforms. The purpose of this study is to elucidate the role of these mutations in the enzyme and also to incorporate unique findings into breast cancer studies. At this point two mutations have been generated pQ293R and pC412S. However, pC412S is still in the process of being overexpressed and purified in order to be tested. The total enzymatic yield for pQ293R was 13.124 mg (6.607 mg/L of media) and had an enzymatic activity of 0.061 U/mg. This value show that the mutated enzyme has less activity than the wildtype, which has an activity value of 0.11 U/mg. Additional work is underway to determine if these mutations affect the ability of the enzyme to incorporate copper or to generate LTQ.