The X-ray structure of human serum ceruloplasmin at 3.1 Å: nature of the copper centres

The X-ray structure of human serum ceruloplasmin has been solved at a resolution of 3.1?Å. The structure reveals that the molecule is comprised of six plastocyanin-type domains arranged in a triangular array. There are six copper atoms; three form a trinuclear cluster sited at the interface of domains 1 and 6, and there are three mononuclear sites in domains 2, 4 and 6. Each of the mononuclear coppers is coordinated to a cysteine and two histidine residues, and those in domains 4 and 6 also coordinate to a methionine residue; in domain 2, the methionine is replaced by a leucine residue which may form van der Waals type contacts with the copper. The trinuclear centre and the mononuclear copper in domain 6 form a cluster essentially the same as that found in ascorbate oxidase, strongly suggesting an oxidase role for ceruloplasmin in the plasma.     

Severe thrombocytopenia associated with acute folic acid deficiency and severe hemorrhage in two patients.

Severe thrombocytopenia associated with acute folic acid deficiency in two inpatients is described. Both patients had severe hemorrhagic manifestations. A similar syndrome has previously been described in patients who have undergone extensive surgery or dialysis or both. The present cases show that acute folic acid deficiency can occur in other circumstances and may go unrecognized because other signs in the peripheral blood of a megaloblastic process may be minimal. A good response was obtained with folic acid therapy.     

Biological activity of gibberellin analogues

In order to determine the significance of the C-6 carboxyl group for the biological activity gibberellin A₃, 6-epigibberellin A₃, 7-norgibberellin A₃, 6-methyl-7-norgibberellin A₃, and 7-homogibberellin A₃ were studied using dwarf pea, dwarf maize, dwarf rice, dwarf barley and -amylase bioassays. All gibberellin A₃(GA₃)derivatives tested were considerably less active than GA₃. In all biossays, 6-epi-GA₃ showed a low activity of the same order, whereas 6-methyl-7-nor-GA₃ was inactive. Surprisingly, 7-nor-GA₃ had some activity in the dwarf rice (root application), dwarf barley, and -amylase bioassay, in contrary to its low potency in the dwarf pea, dwarf maize, and dwarf rice (micro drop) bioassay. 7-Homo-GA₃ was primarily active in the dwarf maize, dwarf barley and dwarf rice bioassay. It also caused antigibberellin effects in dwarf rice. The results demonstrate that the C-6 carboxyl group is not absolutely essential for biological activity of gibberellins. The different activities of 7-nor-GA₃ observed in the various test systems may indicate that the C-6 carboxyl group is a structural requirement more for uptake and/or transport processes than for receptor affinity.Abbreviation GA₃ gibberellic acid     

Enzymatic amplification involving glycosyltransferases forms the basis for the increased size of asparagine-linked glycans at the surface of NIH 3T3 cells expressing the N-ras proto-oncogene.

Expression of ras oncogenes in NIH 3T3 fibroblasts results in the acquisition by these cells of an invasive potential concomitant with the appearance of cell surface asparagine-linked complex-type glycan structures of a higher average molecular weight (Bolscher, J.G. M., van der Bijl, M. M. W., Neefjes, J. J., Hall, A., Smets, L.A., and Ploegh, H.L. (1988) EMBO J. 7, 3361-3368). We have investigated the enzymatic basis for the altered glycosylation by assessing the activities of all major Golgi glycosyltransferases involved in the synthesis of these structures. Use was made of a stable transfectant cell line (T15) containing the N-ras-protooncogene under the control of a glucocorticoid-inducible mouse mammary tumor virus promoter. Upon induction of the ras gene with dexamethasone: 1) the levels of N-acetylglucosaminyltransferase I and II were essentially unaltered, indicating an unaffected potential to synthesize complex-type glycans; 2) the activities of the branching N-acetylglucosaminyltransferase III and V were elevated 2- to 2.5-fold suggesting the formation of increased amounts of bisected glycans and of structures carrying a Gal beta 1----GlcNAc beta 1----6Man-branch; 3) the levels of the elongating beta 4-galactosyltransferase and beta 3-N-acetylglucosaminyl-transferase were increased 5- to 7-fold indicating a strongly enhanced capacity to synthesize polylactosaminoglycan chains; 4) the level of the major chain-terminating enzyme, alpha 3-galactosyltransferase, was slightly decreased (0.7-fold), whereas those of the alpha 3- and alpha 6-sialyltransferases were slightly elevated (1.3- and 2-fold, respectively), suggesting a shift from termination by alpha-galactosyl residues to termination by sialic acid moieties. Studies on the acceptor specificities of the different glycosyltransferases indicate that these changes occur in a coordinated manner in which the effects of altered glycosyltransferase expression levels amplify each other. Analysis of the size of cell surface complex-type glycopeptides before and after digestion with neuraminidase and endo-beta-galactosidase suggested an increased sialic acid density, an increase in the number and/or length of polylactosaminoglycan chains, and an increased branching of the glycans upon N-ras induction. The enzymatic results explain these structural changes and allow us to define the alterations in glycosylation pathways associated with ras expression.     

Analysis of cellular interactions in density-dependent inhibition of 3T3 Cell proliferation

Subpopulations of different proliferative status are determined during cell-density dependent proliferation of 3T3 cells. From these data the probability of conversion of proliferative to quiescent cells is derived and found to correlate well with published data on binding of growth-inhibiting factors secreted from growth-inhibited cells.Based on material presented at the Symposium Intercellular Communication Stuttgart, September 16–17, 1982     

Formation of human immunoreactive LMW and HMW-kininogens during coagulation. Comparison with bradykinin activity

Variations of the levels of human HMW and LMW kininogens have been studied in serum and in plasma incubated in vitro during 30 hours, at three different temperatures: 4 degrees, 22 degrees and 37 degrees C. There is a small difference between the level of LMW kininogen in plasma and in serum, but the serum and plasma level of LMW kininogen are almost stable during the time of incubation at the three temperatures. However, HMW kininogen is reduced in serum to about fifty percent of its plasma level. It decreases in plasma and in serum during incubation, overall at 4 degrees C. A strict parallelism stays between the radioimmunoassay and the bioassay.