Identification of Proteins Differentially Expressed in the Conventional Renal Cell Carcinoma by Proteomic Analysis

Renal cell carcinoma (RCC) is one of the most malignant tumors in urology, and due to its insidious onset patients frequently have advanced disease at the time of clinical presentation. Thus, early detection is crucial in management of RCC. To identify tumor specific proteins of RCC, we employed proteomic analysis. We prepared proteins from conventional RCC and the corresponding normal kidney tissues from seven patients with conventional RCC. The expression of proteins was determined by silver stain after two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The overall protein expression patterns in the RCC and the normal kidney tissues were quite similar except some areas. Of 66 differentially expressed protein spots (p<0.05 by Student t-test), 8 different proteins from 11 spots were identified by MALDI-TOF-MS. The expression of the following proteins was repressed (p<0.05); aminoacylase-1, enoyl-CoA hydratase, aldehyde reductase, tropomyosin alpha-4 chain, agmatinase and ketohexokinase. Two proteins, vimentin and alpha-1 antitrypsin precursor, were dominantly expressed in RCC (p<0.05).

Immunocytochemical localization of amidating enzymes in the sustentacular cells of phaeochromocytoma

To evaluate the presence of amidating enzymes in phaeochromocytoma. Materials and An immunocytochemical study of 21 cases of benign and malignant phaeochromocytomas of the adrenal medulla, was carried out to demonstrate peptidylglycine-alpha- amidating monooxogenase enzymes, using avidin-biotin-peroxidase complex method on paraffin blocks, applying S-100 protein, peptidylgylcine-alpha- hydroxylating monooxogenase and peptidyl-alpha- hydroxyglycine-alpha- amidating lyase as the first layer on paraffin serial sections. Chromaffin cells were positively stained by neuron-specific-enolase, 20/21 were positive for chromogranin, while sustentacular cells were negative. Sustentacular cells were positive for S-100 protein, while phaeochromocytes were negative. All sustentacular cells as well as Schwann cells were positive for S-100 protein and peptidylglycine-alpha- hydroxylating monooxogenase. The stain for the latter peptide was less intense, while chromaffin cells were all negative. Localization of peptidylglcine-alpha- hydroxylating monooxogenase in sustentacular cells suggest that these cells can produce amidating enzymes

Purification and characterization of extracellular penicillin V acylase from Fusarium sp. SKF 235.

Penicillin V acylase from Fusarium sp. SKF 235 culture filtrate was purified to homogeneity. The enzyme was a glycoprotein and composed of single polypeptide chain with molecular weight of 83,200 Daltons. The pH and temperature optima were 6.5 and 55 degrees C, respectively. The KM for penicillin V was 10 mM but the enzyme was inhibited by penicillin V at concentrations above 50 mM. Products of reaction, 6-aminopenicillanic acid and phenoxyacetic acid inhibited the enzyme competitively and noncompetitively with Ki values of 18 mM and 45 mM, respectively. The enzyme specifically hydrolyzed penicillin V, cephalosporanic acid V and penicillin V sulphoxide. Other phenoxy acetyl amides studied were not hydrolysed. It is proposed that phenoxyacetyl moiety alone is not recognized by the penicillin V acylase and in addition, the beta-lactam structure contributes in formation of enzyme-substrate complex.