Vol. 3 No. 1 March 1998

Volume 3 (1998) pp 3-11
Title INCREASED ACTIVITY OF PHOSPHOLIPASE C IN AORTAS OF SPONTANEOUSLY HYPERTENSIVE RATS CORRELATES WITH DECREASED SPHINGOMYELIN : TOTAL PHOSPHOLIPID RATIO
Authors E. Gryckiewicz, A. Dettlaff and T. Pawełczyk*
Abstract The activity of phospholipase C (PLC) in the aortas of spontaneously hypertensive rats (SHR) was higher than in the aortas of age-matched normotensive Wistar-Kyoto rats (WKY). This was associated with the higher level of inositol 1,4,5-trisphosphate and diacylglycerol in aortas of SHR compared to the level of these compounds found in aortas of WKY. Observed changes in PLC activity correlates with changes in phospholipid composition of SHR aortas. The sphingomyelin (SM) to total phospholipid ratio decreased significantly in aortas of SHR compared to WKY. Since, SM was proposed to be the major PLC d inhibitor in vivoit might be possible that observed higher activity of PLC in aortas of SHR results from decreased content of this phospholipid.
Address and Contact Information Department of Clinical Biochemistry, Medical University of Gdańsk, 80-211 Gdańsk, Poland
*To whom correspondence should be adressed. Tel. (058) 347-82-22 ext. 17-76, Fax (058) 344-96-53, e-mail; tkpaw@amedec.amg.gda.pl
[Rozmiar: 1332 bajtów]

Volume 3 (1998) pp 13-20
Title ARSENICAL - INDUCED TRANSCRIPTIONAL ACTIVATION OF THE YEAST Saccharomyces cerevisiae ACR2 AND ACR3 GENES REQUIRES THE PRESENCE OF THE ACR1 GENE PRODUCT
Authors P. Bobrowicz and S. Ułaszewski*1
Abstract The yeast ACR1 gene encodes a putative transcriptional regulatory bZIP protein involved in the arsenical resistance. We demonstrate that the ACR2 and ACR3 genes are positively regulated by Acr1p. Changes in the ACR1 gene dosage influence the ACR2-lacZ and ACR3-lacZ fusion genes response to the arsenite induction. Arsenic and antimony compounds, but not bismuth and cadmium salts, strongly induced the expression of the ACR3 gene encoding the arsenite transporter related to the prokaryotic ArsB proteins.
Address and Contact Information Institute of Microbiology, Wroclaw University, Przybyszewskiego 63, 51-148 Wroclaw, Poland
* To whom correspondence should be addressed. TEL: 48-71-3247293; FAX: 48-71-3252151; E-mail: stan@microb.uni.wroc.pl
[Rozmiar: 1332 bajtów]

Volume 3 (1998) pp 21-24
Title EASY PCR SCREENING OF Pichia pastoris TRANSFORMANTS
Authors D. Arora, A. Chauhan and N. Khanna*
Abstract A simple protocol for easy PCR screening of P. pastoris transformants is described. In short, the P. pastoris cells are lysed with very small amount of the enzyme Zymolase and the crude cell lysate is directly used in PCR. This protocol needs no tube transfer steps and also obliviates the requirement of freezing the samples at -800C before PCR screening. Because of a single step screenig, both overall and actual hands on time are considerably reduced.
Address and Contact Information 1 International Centre for Genetic Engineering and Biotechnology, P.O. Box 10504, Aruna Asaf Ali Marg, New Delhi - 110 067, India
*To whom correspondence should be addressed
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Volume 3 (1998) pp 25-35
Title IMMUNOSPECIFIC PROTEIN WITH MOL. WT OF 38/39 kDa FROM LYMPHOCYTIC LEUKEMIC CELLS
Authors Z. M. Kiliańska*1, A. Ptasińska1, J. Błoński2, J. Chruściel1, P. Szymczyk1, E. Krykowski2 and T. Robak2.
Abstract One dimensional patterns of proteins from homogenates and four cellular fractions i.e., nuclear, mitochondrial, microsomal, and cytosolic from normal and leukemic lymphocytes were compared. Results obtained revealed that the neoplastic transformation of normal lymphocytes into CLL and ALL ones is associated with the expression of some novel proteins. Electrophoretically-specific nuclear protein of B-CLL lymphocytes with mol. wt of 38/39 kDa was used as immunogen to produce rabbit antiserum. It was observed that obtained antiserum crossreacted with 38/39 kDa antigen of nuclear fractions from CLL and ALL lymphocytes (15 of 16 studied), but not with any of normal ones. It was shown, by Western blot technique, that the expression of 38/39 kDa antigen is correlated with progression of B-CLL disease.
Address and Contact Information 1Department of Cytobiochemistry, University of Łódź, Banacha 12/16, 90-237 Łódź, Poland,
2Department of Haematology, Medical University of Łódź, Paderewskiego 4, 93-513 Łódź, Poland.
*Reprint address
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Volume 3 (1998) pp 37-48
Title PURIFICATION OF THE Ca2+ - ATPase FROM RAT CORTICAL, CEREBELLAR AND HIPPOCAMPAL SYNAPTOSOMAL MEMBRANES BY TWO - STEP CHROMATOGRAPHY
Authors L. Żylińska*, L. Lachowicz and E. Gromadzińska
Abstract Ca2+-ATPase, the enzyme responsible for maintenance of low resting Ca2+ level in the nerve cell, has been purified from rat cortical, cerebellar and hippocampal synaptosomal membranes by affinity chromatography on Calmodulin-Agarose which followed Reactive Red 120-Agarose column. The enzymes from these three regions ran as broad, monomeric bands on SDS-PAGE with a molecular weight at range 130 -138 kDa, and were identified as a Ca2+-ATPase using monoclonal antibody 5F10. Analysis of the kinetic parameters revealed that hippocampal Ca2+-ATPase exhibited 2 times higher affinity for ATP, than cortical and cerebellar enzymes. The affinity for Ca2+ increased in order: cerebellum, cortex, and hippocampus. The differences in kinetic characteristics of purified enzymes, suggest that in adult rat brain the Ca2+-ATPase could be represented at a protein level by the region-dependent combination of several isoforms.
Address and Contact Information II Department of Biochemistry, Medical University of Łódź,6 Lindley Street, 90-131 Łódź, Poland
*Corresponding author
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Volume 3 (1998) pp 49-56
Title PLASMINOGEN ACTIVATOR INHIBITOR-1 (PAI-1) GENE 4G/5G PROMOTER POLYMORPHISM IN SUBJECTS WITH CANCER
Authors B. Smolarz, J. Błasiak, D. Piestrzeniewicz and J. Pytel
Abstract Plasminogen activator inhibitor PAI-1 is a fast-acting controlling factor of the plasminogen activator system. The system contains proteolytic enzymes that may contribute to cancer cell invasion by degrading the surrounding extracellular matrix and dissociate cell-cell or cell-matrix attachments. There is substantial evidence in many types of cancer that the antigen content of PAI-1 in primary cancer tissue extracts are of strong prognostic value: high level of PAI-1 in tumor predict poor prognosis for the patient. Moreover, it was demonstrated that the level of PAI-1 in metastasis is significantly higher compared to primary tumors. An insertion/deletion polymorphism in the promoter of the PAI-1 gene has been described that relates to plasma PAI-1 levels. We studied this polymorphism in the plasma of subjects with cancer. Blood was taken from 23 patients (10 breast cancers, 5 gastric cancers, 4 head and neck cancers, 2 melanomas and 2 colorectal cancers) and 23 matched controls. Although frequencies of the 4G and 5G alleles were approximately 0.5/0.5 in both patients and controls, the genotype distribution differed significantly between the two groups – the 4G/5G genotype was observed more frequently in patients with cancer than in the controls. Further studies are needed to check whether the prevalence of the 4G/5G genotype may influence an individual’s plasminogen activation system capacity and thereby contribute in a small way to the cancer risk profile.
Address and Contact Information University of Łódź, Department of Molecular Genetics, Łódź, Poland and Medical Academy of Łódź, Department of Oncology, Łódź, Poland Correspondence to: Janusz Blasiak, University of Łódź, Department of Molecular Genetics, ul. Banacha 12/16, 90-237 Łódź, Poland. FAX +48-42 35 44 84, e-mail januszb@biol.uni.lodz.pl
[Rozmiar: 1332 bajtów]

Volume 3 (1998) pp 57-73
Title MODULATION OF PLANT NUCLEAR MATRIX NUCLEOLYTIC ACTIVITY AND SPECIFICITY MAY RESULT FROM INTERACTION OF NUCLEASE-PROTEIN 14-3-3 COMPLEX WITH INNER NUCLEAR MEMBRANE LIPIDS
Authors J. Lubocka1 and A. Kozubek2*
Abstract Plant cell nuclear matrix depleted in lipids exhibited approximately 50% lower endonucleolytic activity attributed to 32 kDa nuclease. Supplementation of delipidated matrix with phospholipids, sterols and glycolipids resulted in the recovery of nuclease activity. The extent of recovery is dependent upon the type of lipid used for reconstitution. The most effective (over 97%) in recovery of the nucleolytic activity were PC, PE, DGDG and stigmasterol. Some recovery is also observed when other natural amphiphilic molecules are studied (resorcinolic lipids). Recombinant plant protein 14-3-3 showed ability for direct interaction with lipid monolayers. The interaction was confirmed by analysis of the monolayer collected from the subphase after incubation with protein 14-3-3. Deletion of the 12 kDa N-terminal fragment of the protein 14-3-3 abolished its ability for binding to lipid monolayer. The results suggest that in vivo direct interaction of protein 14-3-3 with nuclear envelope lipids may participate in modulation of the nuclease 32 kDa activity. A postulated model of the interactions is discussed.
Address and Contact Information 1Department of Genetic Biochemistry,
2Department of Lipids and Liposomes
Institute of Biochemistry and Molecular Biology, University of Wroclaw Przybyszewskiego 63/77, 51-148 Wroclaw, Poland
*Corresponding author: tel/fax: (48) +71 3252930, E-mail: kozubek@microb.uni.wroc.pl
[Rozmiar: 1332 bajtów]


Volume 3 (1998) pp 75-90
Title MANIPULATING OF 14-3-3 PROTEIN EXPRESSION RESULTES IN THE CHANGES OF CATECHOLAMINE CONTENT IN POTATO PLANT
Authors G. Wilczynski, A. Kulma, I. Feiga, A. Wenczel* and J. Szopa**
Abstract Recently transgenic potato plants were created where the 14-3-3 protein derived from Cucurbita pepo was overexpressed or potato endogenous 14-3-3 protein repressed. Detailed analysis of those plants suggested that a isolated 14-3-3 isoforms are involved in the control of plant senescence and carbohydrate metabolism. In this study carbohydrate content, adenine nucleotide level and catecholamine content in tubers and leaves of transgenic plants grown in greenhouse and in a field were compared. Overexpressing of 14-3-3 protein gave rise to increase catecholamine contents and soluble sugars in leaves and a reduction in tubers size and starch content. The repression of 14-3-3 synthesis led to opposite effect, a decrease in catecholamine, soluble sugars contents in leaves, and an increase in tubers size and starch content. It is proposed that 14-3-3 protein affects carbohydrate metabolism in potato via regulation of catecholamine synthesis.
Address and Contact Information Institute of Biochemistry, University of Wroclaw, Przybyszewskiego 63/77 51-148 Wroclaw, Poland
*Zentrallaboratorium im Univ.-Klinikum Erlangen, Krankenhausstr.12, 91054 Erlangen, Germany
**Corresponding author, E-mail: szopa@microb.uni.wroc.pl
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