| Autor: |
Peitsaro, Nina, Polianskyte, Zydrune, Eriksson, Ove |
| Předmět: |
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| Zdroj: |
FASEB Journal; Apr2007, Vol. 21 Issue 5, pA669-A669, 1/5p |
| Abstrakt: |
The mammalian active site serine enzyme LACTB has evolved from bacterial penicillin-binding proteins. Orthologs of LACTB are found in all vertebrates. To gain insight into the physiological function of LACTB we have investigated the expression level and intracellular localization of LACTB. We found that LACTB is ubiquitously expressed in human and rodent tissues. However, the expression level correlated with the metabolic activity of the tissue suggesting that LACTB is involved in mitochondrial housekeeping. To investigate the intracellular distribution of LACTB we cloned the human LACTB coding sequence into the pcDNA-DEST47 plasmid resulting in a LACTB-GFP construct. Transfection of human breast cancer MCF-7 cells with the LACTB-GFP construct showed that LACTB is targeted solely to mitochondria. Furthermore, immunoelectron microscopy indicated that LACTB is located in the intermembrane space. To study LACTB in a loss of function model we made six different constructs for RNA silencing. The constructs were cloned into a pLL3.7 plasmid for lentivirus delivery into MCF-7 cells. Based on real-time PCR all constructs decreased the LACTB mRNA level significantly. Our results show that LACTB is a ubiquitous mitochondrial protein localized in the intermembrane space and that the siRNA technique is feasible for further investigations into the cell biological function of LACTB. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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