Search results

1

Biosynthesis and distribution of leucocyte elastase inhibitor. Production of recombinant inhibitor

100%

Kasza A., Korpula-Mastalerz R., Rose-John S., Dubin A.

Acta Biochimica Polonica

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1996

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vol. 43

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issue 3

497-501

2

Evaluation of P1' substrate specificity of staphylococcal SplB protease

88%

Pustelny K., Stach N., Wladyka B., Dubin A., Dubin G.

|

EN

Staphylococcus aureus is a dangerous human pathogen characterized by growing antibiotic resistance. Virulence of S. aureus relies on a variety of secreted and cell surface associated virulence factors among which certain proteolytic enzymes play an important role. Amid staphylococcal extracellular proteases, those encoded by the spl operon remain poorly characterized, both in terms of enzymology and their physiological role. Initial data demonstrated that Spl proteases exhibit restricted substrate specificity. This study describes development of convenient protein FRET substrates for SplB protease and characterization of the substrate preference of the protease at the P1' position. Kinetic data on hydrolysis of a panel of substrates substituted at the said position is provided.

3

A systematic investigation of the stability of green fluorescent protein fusion proteins

76%

Janczak M., Bukowski M., Górecki A., Dubin G., Dubin A., Wladyka B.

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2015

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vol. 62

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issue 3

407-411

EN

X-ray crystallography provides important insights into structure-function relationship in biomolecules. However, protein crystals are usually hard to obtain which hinders our understanding of multiple important processes. Crystallization requires large amount of protein sample, whereas recombinant proteins are often unstable or insoluble. Green fluorescent protein (GFP) fusion is one of the approaches to increase protein synthesis, solubility and stability, facilitating crystallization. In this study we analyze the influence of the linker length, composition and the position of GFP relative to the fusion partner on the fusion protein production and stability. To this end, multiple constructs of enzymatically impaired variant of PemKSa toxin from Staphylococcus aureus CH91 fused to GFP were generated. Fusion protein production in Escherichia coli was evaluated. The proteins were purified and their stability tested. PemKSa-α14aa-GFP fusion provided best production and stability. Obtained results demonstrate the importance of optimization of fusion protein construct, including linker selection and the order of fusion partners, in obtaining high quantities of stable protein for crystallization.

4

Comparative analysis of RCAS1 level in neoplasms and placenta.

76%

Wicherek L., Dutsch M., Mak P., Klimek M., Skladzien J., Dubin A.

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2003

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vol. 50

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issue 4

1187-1194

EN

The tumor associated antigen RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) expressed with high frequency in various cancer and trophoblast cells, inhibits growth of estrogen receptor-expressing cells and induces apoptosis. Because previous reports demonstrated RCAS1 presence only by non-quantitative immunocytochemistry methods, we decided to use a Western blotting with anti-RCAS1 monoclonal antibodies for estimation of the relative content of the tumor-associated antigen. One hundred tissue samples were assayed (neoplasms, chronic inflammatory diseases, healthy tissues, trophoblasts and placentas at term). RCAS1 was present in all neoplastic, placental and trophoblast tissue samples and its level in malignant samples was statistically significantly higher than in benign neoplasms. The amount of RCAS1 in chronic inflammations was also significantly increased in immune mediated diseases, like allergic nasal polyps and sarcoidosis. The RCAS1 protein was not revealed in healthy mucous membrane and in muscle tissues. The presented results suggest that RCAS1 might play an important role in tumor escape from host immunological surveillance and carry weight in the down regulation of the maternal immune response, thereby maintaining pregnancy.

5

A novel member of the thermolysin family, cloning and biochemical characterization of metalloprotease from Staphylococcus pseudintermedius

76%

Wladyka B., Bista M., Sabat A. J., Bonar E., Grzeszczuk S., Hryniewicz W., Dubin A.

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2008

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vol. 55

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issue 3

525-536

EN

Thermolysins constitute a family of secreted bacterial metalloproteases expressed, among others, by several pathogens. Strains of Staphylococcus pseudintermedius isolated from diseased dogs and judged as protease-positive, by skim milk agar plate culture, were investigated for protease content. No proteolytic activity was detected when the bacteria were grown in regular liquid media. Unexpectedly, supplementation of the medium with calcium ions resulted in expression of a metalloprotease and profound changes in the profile of extracellular proteins. On the basis of homology to other staphylococcal metalloproteases, the nucleotide sequence of the gene encoding this protease (Pst) and its flanking regions was determined. The full-length pst codes for a protein with an open reading frame of 505 amino acids. The internal region contains the HEXXH catalytic domain that is conserved in members of the thermolysin family. Regardless of the presence of calcium in the medium, the expression of the protease gene was of the same intensity. This suggests that regulation of the metalloprotease production by calcium ions is at a post-transcriptional level. Isolates of S. pseudintermedius exhibit a proteolytic phenotype due to the metalloprotease expression, however only in presence of calcium ions, which most probably stabilize the structure of the protease.

6

Evidence for the presence of different alpha-1-proteinase inhibitor genes products in mouse plasma

75%

Dubin A., Mak P., Travis J.

Acta Biochimica Polonica

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1996

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vol. 43

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issue 3

475-479

7

New generation of peptide antibiotics

64%

Dubin A., Mak P., Dubin G., Rzychon M., Stec-Niemczyk J., Wladyka B., Maziarka K., Chmiel D.

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2005

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vol. 52

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issue 3

633-638

EN

The increasing antibiotic resistance of pathogenic bacteria calls for the development of alternative antimicrobial strategies. Possible approaches include the development of novel, broad-spectrum antibiotics as well as specific targeting of individual bacterial virulence factors. It is impossible to decide currently which strategy will prove more successful in the future since they both promise different advantages, but also introduce diverse problems. Considering both approaches, our laboratory's research focuses on the evaluation of hemocidins, broad-spectrum antibacterial peptides derived from hemoglobin and myoglobin, and staphostatins, specific inhibitors of staphopains - Staphylococcus aureus secreted proteases that are virulence factors regarded as possible targets for therapy. The article summarizes recent advances in both fields of study and presents perspectives for further development and possible applications.

8

The intracellular serpin family

50%

Korpula-Mastalerz R., Dubin A.

Acta Biochimica Polonica

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1996

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vol. 43

|

issue 3

419-429

Refine search results

Biosynthesis and distribution of leucocyte elastase inhibitor. Production of recombinant inhibitor

Evaluation of P1' substrate specificity of staphylococcal SplB protease

A systematic investigation of the stability of green fluorescent protein fusion proteins

Comparative analysis of RCAS1 level in neoplasms and placenta.

A novel member of the thermolysin family, cloning and biochemical characterization of metalloprotease from Staphylococcus pseudintermedius

Evidence for the presence of different alpha-1-proteinase inhibitor genes products in mouse plasma

New generation of peptide antibiotics

The intracellular serpin family