Susceptibility of neuroactive peptides to aminopeptidase digestion is related to molecular size.

Peptide fragments derived from the NH₂-terminus of corticotropin were found to exhibit widely differing degrees of stability to degradation by aminopeptidase M. Corticotropin itself was 135 times more stable than its NH₂-terminal pentapeptide, and similar differences in stability were observed with peptides derived from the B-chain of bovine insulin. Enkephalin linked covalently to the A-chain of bovine insulin was at least 100 times more stable than the pentapeptide. The results demonstrate that the molecular size of a peptide is one factor that determines its NH₂-terminal stability.     

Biochemical, biological, and immunological properties of chemically deglycosylated human choriogonadotropin

A chemical method of deglycosylation of human choriogonadotropin (hCG) was used to assess the role of carbohydrate moiety in the maintenance of quaternary structure and functional parameters such as receptor binding, immunological activity, and in vitro biological response. Treatment of purified hCG with anhydrous HF at 0 degrees C for 60 min was effective in removing more than 75% of the carbohydrate moiety. This extent of deglycosylation altered its chromatographic characteristics as revealed by retarded behavior on Sephadex G-100 and failure to be retained on concanavalin A-Sepharose. The electrophoretic heterogeneity present in native hCG was markedly reduced by deglycosylation. The deglycosylated hCG was stable in the lyophilized form and retained its quaternary structure as revealed by the fluorescence probe 8-anilino 1-naphthalene sulfonic acid, receptor binding, and immunological activities. Unlike receptor binding and immunological activities, which were fully retained, the ability of the hormone to stimulate cyclic AMP accumulation in vitro in rat interstitial cells was completely abolished.     

Prolonged alteration in composition of fast transported protein in axons prevented from regenerating after injury

A previous study revealed a characteristic alteration in the ratio of labeling of two fast axonally transported polypeptides identified as S1 (22,000 daltons) and S2 (18,000 daltons) and their ratio (S2/S1) in rat motoneuron axons following axonal injury and subsequent regeneration. In this study the S2/S1 ratio was determined for axons which were resected and ligated proximally and which did not regenerate to reinnervate muscle. While the initial increase in the S2/S1 ratio following section was the same as that following a crush, the S2/S1 ratio did not return towards normal values after 42 days, but remained elevated for at least 98 days after injury. It is concluded that the return of S2/S1 ratio to normal values, like some other manifestations of the cell body reaction, is delayed if the injured axons do not regenerate.     

Glycoproteins of trypanosomes: their biosynthesis and biological significance

1. Trypanosomes are unicellular parasites that cause human sleeping sickness in Africa and Chagas' disease in South America. Glycoproteins are important components of their plasma membrane. 2. The bloodstream form of the extracellular salivarian African trypanosome (e.g. Trypanosoma brucei) has the ability to express on its cell surface a repertoire of variant surface glycoproteins (VSGs) and in so doing, evades the immune response of the host (antigenic variation). 3. The VSG is probably synthesized initially in a manner like that of the membrane-bound glycoproteins of mammalian systems, but it also undergoes some novel post-translational modifications. 4. The stercorarian South American trypanosome (Trypanosoma cruzi) is an intracellular parasite which expresses different glycoproteins on its plasma membrane at various stages of its life-cycle, but does not exhibit antigenic variation. 5. The biosynthesis and functions of trypanosomal glycoproteins are compared with those of mammalian glycoproteins, and are discussed with particular reference to potential targets for chemotherapy and immunotherapy of trypanosomiasis.     

Flavin-dependent thymidylate synthase: A novel pathway towards thymine

For several decades only one chemical pathway was known for the de novo biosynthesis of the essential DNA nucleotide, thymidylate. This reaction catalyzed by thyA or TYMS encoded thymidylate synthases is the last committed step in the biosynthesis of thymidylate and proceeds via the reductive methylation of uridylate. However, many microorganisms have recently been shown to produce a novel, flavin-dependent thymidylate synthase encoded by the thyX gene. Preliminary structural and mechanistic studies have shown substantial differences between these deoxyuridylate-methylating enzymes. Recently, both the chemical and kinetic mechanisms of FDTS have provided further insight into the distinctions between thyA and thyX encoded thymidylate synthases. Since FDTSs are found in several severe human pathogens their unusual mechanism offers a promising future for the development of antibiotic and antiviral drugs with little effect on human thymidylate biosynthesis.