| 1. |
- Aaltonen, Emil
(författare)
-
Prokaryotic Arsenic Resistance - Studies in Bacillus subtilis
- 2008
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Arsenic is a toxic metalloid which is found all over the globe. Due to its toxicity and wide abundance, all living organisms have evolved intrinsic arsenic resistance systems. In this study, three proteins that provide arsenic resistance in the Gram-positive bacterium Bacillus subtilis have been investigated. Acr3 is a trans-membrane protein that extrudes arsenite to the cell exterior. It belongs to the ACR family of arsenite transporters. Experimental data on the topology of Acr3, the first ever for a member of the ACR family, is provided and show that Acr3 has 10 trans-membrane helices. Both the N- and C-terminal ends of Acr3 are located to the cytoplasm and the protein has unusually short loops connecting its helices. ArsR is an arsenite sensitive transcription regulator that controls the expression of genes encoding arsenic resistance proteins. Experiments show that the operator site for ArsR from B. subtilis consists of a 6-6-6 inverted repeat and that DNA binding by ArsR involves formation of higher order multimers of the protein. The ArsK (former YqcK) protein has an unknown function. The present study shows that the arsK gene contributes to resistance towards both arsenite and arsenate. The results indicate that the function of ArsK is important in an aerobic environment and that it decreases the inhibitory effect that arsenite has on the sporulation efficiency of B. subtilis. A function of ArsK that involves an enzymatic addition of low molecular weight thiols to arsenic is proposed.
|
|
| 2. |
- Aaltonen, Emil, et al.
(författare)
-
Transmembrane topology of the Acr3 family arsenite transporter from Bacillus subtilis.
- 2008
-
Ingår i: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 0005-2736. ; 1778:4, s. 963-973
-
Tidskriftsartikel (refereegranskat)abstract
- The transmembrane topology of the Acr3 family arsenite transporter Acr3 from Bacillus subtilis was analysed experimentally using translational fusions with alkaline phosphatase and green fluorescent protein and in silico by topology modelling. Initial topology prediction resulted in two models with 9 and 10 TM helices respectively. 32 fusion constructs were made between truncated forms of acr3 and the reporter genes at 17 different sites throughout the acr3 sequence to discriminate between these models. Nine strong reporter protein signals provided information about the majority of the locations of the cytoplasmic and extracellular loops of Acr3 and showed that both the N- and the C-termini are located in the cytoplasm. Two ambiguous data points indicated the possibility of an alternative 8 helix topology. This possibility was investigated using another 10 fusion variants, but no experimental support for the 8 TM topology was obtained. We therefore conclude that Acr3 has 10 transmembrane helices. Overall, the loops which connect the membrane spanning segments are short, with cytoplasmic loops being somewhat longer than the extracellular loops. The study provides the first ever experimentally derived structural information on a protein of the Acr3 family which constitutes one of the largest classes of arsenite transporters.
|
|
| 3. |
- Eriksson, Per-Olof, et al.
(författare)
-
Novel Nano-Sized MR Contrast Agent Mediates Strong Tumor Contrast Enhancement in an Oncogene-Driven Breast Cancer Model.
- 2014
-
Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 9:10
-
Tidskriftsartikel (refereegranskat)abstract
- The current study was carried out to test the potential of a new nanomaterial (Spago Pix) as a macromolecular magnetic MR contrast agent for tumor detection and to verify the presence of nanomaterial in tumor tissue. Spago Pix, synthesized by Spago Nanomedical AB, is a nanomaterial with a globular shape, an average hydrodynamic diameter of 5 nm, and a relaxivity (r1) of approximately 30 (mM Mn)-1 s-1 (60 MHz). The material consists of an organophosphosilane hydrogel with strongly chelated manganese (II) ions and a covalently attached PEG surface layer. In vivo MRI of the MMTV-PyMT breast cancer model was performed on a 3 T clinical scanner. Tissues were thereafter analyzed for manganese and silicon content using inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The presence of nanomaterial in tumor and muscle tissue was assessed using an anti-PEG monoclonal antibody. MR imaging of tumor-bearing mice (n = 7) showed a contrast enhancement factor of 1.8 (tumor versus muscle) at 30 minutes post-administration. Contrast was retained and further increased 2-4 hours after administration. ICP-AES and immunohistochemistry confirmed selective accumulation of nanomaterial in tumor tissue. A blood pharmacokinetics analysis showed that the concentration of Spago Pix gradually decreased over the first hour, which was in good agreement with the time frame in which the accumulation in tumor occurred. In summary, we demonstrate that Spago Pix selectively enhances MR tumor contrast in a clinically relevant animal model. Based on the generally higher vascular leakiness in malignant compared to benign tissue lesions, Spago Pix has the potential to significantly improve cancer diagnosis and characterization by MRI.
|
|
