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Biomedical subjects

E R Podack

Publications and source records attributed to E R Podack.

161 records · Page 9Linked to original sources
PubMed1977

The SC5b-7 complex: formation, isolation, properties, and subunit composition.

Activation of C in C8-depleted serum results in the formation of a soluble complex containing C5, C6, and C7. The complex has an electrophoretic mobility of an alpha-globulin, an s-rate of 18.5S, and a m.w. of 668,000 daltons. This complex was isolated and upon SDS polyacrylamide gel electrophoresis it was found to contain, in addition to C5b, C6 and C7, an 88,000 dalton glycoprotein. The protein was identified as the band V protein of the soluble C5b-9 complex. It is referred to as SIIIs-protein, or S-protein. Since the S-protein does not bind to C5b-6, it is concluded that it is incorporated during the fusion of C5b-6 with C7. The SC5b-7 complex exhibits the same neoantigen as the SC5b-9 complex, but compared to the C5b-6 complex it appears to contain an additionally qualitatively distinct neoantigen.

Animals↗
PubMed1976

Purification of the sixth and seventh component of human complement without loss of hemolytic activity.

Procedures for the isolation of the human complement proteins C6 and C7 have been described. These procedures allow isolation of the two proteins without any loss of hemolytic activity. Apparent activity gains of 160% and 140% were observed for C6 and C7, respectively, when the activity of the isolated proteins was compared with their activity in serum. The recovery of C6 was 3.5 to 11% and that of C7 was 7 to 13% of the amount present in serum. C6 has a m.w.of 128,000 and an electrophoretic mobility at pH 8.6 of -2.6 times 10(-5) cm2 s-1 v-1. C7 has a m.w. of 121,000 and an identical electrophoretic mobility. With 3 times 10(7) assay cells, 63% hemolysis was achieved with 1 ng of C6 and 3.8 ng C7. On polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and after reduction with mercaptoethanol, C6 and C7 behaved as single polypeptide chain proteins.

Complement C6↗
PubMed1976

The C5b-9 complex: subunit composition of the classical and alternative pathway-generated complex.

The membrane attack complex of complement (C5b-9) is identical in composition regardless of which pathway of activation was instrumental in its formation. Band V protein was consistently a subunit of the soluble complex. Since band V protein is not required for complement-dependent cytolysis, it probably represents a membrane site equivalent in serum of the nascent C5b-9 complex.

Centrifugation, Density Gradient↗
PubMedDate not supplied

Assembly of two types of tubules with putative cytolytic function by cloned natural killer cells.

The formation of ultrastructural membrane lesions of varying size during cell mediated cytolysis effected by human peripheral blood leukocytes was recently reported by Dourmashkin et al. and Henkart et al. Using cloned mouse natural killer (NK) cells as effectors and YAC-1 cells or rabbit erythrocytes as targets, we now report two types of membrane lesions with inner diameters of 16 +/- 2 nm and approximately 5 nm, respectively. These lesions arise by membrane insertion of tubular complexes that may be assembled from subunits during the cytolytic reaction. The tubules are detected on target membranes by immune electron microscopy and appear to form transmembrane channels as seen in ultrathin sections. Both tubules are partially purified by membrane extraction with SDS and gel filtration in deoxycholate containing buffer. Based on the correlation of tubule assembly and cytolysis and on their detection on target membranes, we suggest that both types of tubules may be related to cytolysis.

Cell Membrane↗
PubMedDate not supplied

Detection of perforin in human peritoneal fluid T-lymphocytes.

OBJECTIVE: Perforin is a specific marker of functionally active cytotoxic T-lymphocytes (CTLs) and natural killer (NK) cells. The purpose of this study was to detect perforin-positive lymphocytes in ascites secondary to various gynecologic malignancies and nonneoplastic conditions. STUDY DESIGN: Fifty-three unselected peritoneal fluid specimens submitted for cytopathologic diagnosis were used. A monoclonal antibody to human perforin was used to examine its expression in mononuclear cells from ascites specimens using a standard immunoperoxidase technique. RESULTS: Strong perforin expression by 15-30% of mononuclear cells was detected in 10 of the 13 patients with severe alcoholic hepatitis, 3 of the 5 patients with chronic active hepatitis without cirrhosis and 1 patient with an autoimmune disorder of unknown etiology. The lymphocytes showed variable positivity for T cell markers (CD2, CD4, CD8 and CD56). Perforin was not detected in ascites specimens obtained from patients with such nonneoplastic disorders as cirrhosis or end stage renal or cardiac failure. Similarly, ascites specimens from patients with various gynecologic malignancies were negative for perforin-positive lymphocytes. CONCLUSION: The detection of cytotoxic lymphocytes in peritoneal fluid obtained from patients with liver injury may have implications for the pathogenesis of this disease.

Antibodies, Monoclonal↗