H.W. Yeung

Proteins with abortifacient, ribosome inactivating, immunomodulatory, antitumor and anti-AIDS activities from Cucurbitaceae plants.

1. The biochemical characteristics and biological activities of eight Cucurbitaceae plant proteins designated trichosanthin (isolated from tubers of Trichosanthes kirilowii), beta-trichosanthin (isolated from tubers of Trichosanthes cucumeroides), alpha- and beta-momorcharins (isolated from seeds of Momordica charantia), momorchochin (isolated from tubers of Momordica cochinchinensis), luffaculin (isolated from seeds of Luffa acutangula) and luffin-a and luffin-b (isolated from seeds of Luffa cylindrica), were reviewed. 2. The isolation procedures for all eight proteins are based on aqueous extraction, acetone fractionation and ion exchange chromatography. Ammonium sulfate precipitation and gel filtration are steps which may be included to improve purification. 3. The proteins are basic in nature and possess a molecular weight of approx. 30,000. All except trichosanthin are glycoproteins. The content of Asx and Glx residues is high. The N-terminal amino acid residue is Asp. Their amino acid compositions and N-terminal amino acid sequences are similar. 4. Circular dichroism spectroscopic studies revealed that trichosanthin, alpha- and beta-momorcharins possess similar secondary but different tertiary structures. 5. Most of the proteins are immunologically distinct. 6. The proteins exhibit abortifacient, antitumor, ribosome inactivating and immunomodulatory activities. Trichosanthin manifests anti-human immunodeficiency virus activity.

Anti-human immunodeficiency virus (anti-HIV) natural products with special emphasis on HIV reverse transcriptase inhibitors

This review article aims at summarizing research findings concerning natural products which are endowed with the ability to inhibit human immunodeficiency virus (HIV). An emphasis is placed on HIV reverse transcriptase inhibitors because the bulk of the literature is focused on these compounds. It was found that a spectacular diversity of chemical structures encompassing proteins, terpenoids, coumarins, xanthones, alkaloids, flavonoids, polyphenols, and polysaccharides, which are elaborated by plant species as phylogenetically remote as the algae, gymnosperms and angiosperms, were capable of rendering the retroviral enzyme less active. The literature pertaining to natural products with HIV protease and integrase inhibitory activities is less voluminous.

A comparison of human immunodeficiency virus type 1 inhibition by partially purified aqueous extracts of chinese medicinal herbs

A multiple screening approach to detect compounds inhibitory to various aspects of the human immunodeficiency virus type 1 (HIV-1) life-cycle has been applied to aqueous extracts of 19 herbs traditionally used in Chinese medicine as anti-viral agents. The extracts were tested for their ability to inhibit HIV-1 in a series of in vitro assays. The extracts were tested for inhibition of the interaction between HIV-1 gp120 and immobilized CD4 receptor, inhibition of recombinant HIV-1 reverse transcriptase and for inhibition of three glycohydrolase enzymes that contribute to viral protein glycosylation. Six of the herb extracts (30%) were potent inhibitors of the interaction between HIV-1 gp120 and the CD4 receptor (ID50 5.6 - 79.4 microg/ml), two extracts (10%) contained potent reverse transcriptase inhibitors (ID50 16.9 - 26.0 microg/ml) and 14 extracts (75%) were able to inhibit at least one of the glycohydrolase enzymes.

Insulin-like molecules in Momordica charantia seeds.

Decorticated Momordica charantia seeds were extracted and processed by a method which was developed originally for the purification of insect and annelid insulins. Essentially, the method entailed HCl--ethanol extraction, neutralization with NH4OH, gel filtration on Sephadex G-50, ion exchange chromatography on CM Sepharose CL-6B and desalting on Sephadex G-10. Of the seven fractions collected, three fractions were obtained with antilipolytic and lipogenic activities in isolated adipocytes and one fraction with only lipogenic activity. The data indicate that molecules with insulin-like bioactivity are present in Momordica charantia seeds.

Minireview: trichosanthin--a protein with multiple pharmacological properties.

Trichosanthin (TCS), a protein isolated from the root tuber of the Chinese medicinal herb Trichosanthes kirilowii Maxim., is used to induce abortion in China. It also possesses immunomodulatory, anti-tumor and anti-human immunodeficiency virus properties. TCS is a member of the family of ribosome-inactivating proteins and inactivates eukaryotic ribosomes via its N-glycosidase activity. The gene encoding TCS has been cloned and over-expressed and the crystal structure of this protein resolved to 1.73A. In this review, the various pharmacological properties of TCS are discussed and assessed.

Demonstration of ribonuclease activity in the plant ribosome-inactivating proteins alpha- and beta-momorcharins.

Alpha- and beta-momorcharins, ribosome-inactivating proteins from Momordica charantia seeds, were utilized in this investigation. Ribonucleolytic cleavage was observed after naked rRNA was incubated with either momorcharin. Beta-momorcharin, and to a lesser extent alpha-momorcharin, also acted on tRNA to release acid-soluble UV-absorbing products. Such activity was optimal at pH around 5.5. Using polyhomoribonucleotides as substrate, it was found that the momorcharins preferentially acted on polyU, but exerted negligible effects on polyA, polyC and polyG. Chromatographic analysis of the reaction product indicated that mono and/or oligo-ribonucleotides, but not free base, were generated from polyU, suggesting that the enzymatic action involved ribonucleolytic cleavage. Similar to the results obtained with tRNA as substrate, beta-momorcharin was about 15-fold more active than alpha-momorcharin on polyU.

Use of trichosanthin to reduce infection by turnip mosaic virus

Abstract Exogenous application of recombinant trichosanthin (TCS) inhibited local lesion formation by turnip mosaic virus (TuMV) in leaves of Nicotiana tabacum and caused a delay in the development of mosaic symptoms by TuMV in Brassica parachinensis . A dose-response relationship was observed in the latter case. To produce transgenic plants expressing a TCS cDNA, a chimeric gene which contains a fusion between the cauliflower mosaic virus 35S promoter, the TCS-encoding cDNA and the octopine synthase 3′ polyadenylation sequence ( ocs 3′) was constructed. The chimeric gene was then cloned into a binary vector and introduced into the genome of N. tabacum cv. ‘Wisconsin 38’ via Agrobacterium tumefaciens -mediated transformation. Three transgenic tobacco plants which synthesized TCS protein in the cytosol were completely resistant to mechanical inoculation of TuMV.

Immunomodulatory effect of a polysaccharide-enriched preparation of Codonopsis pilosula roots

1. A polysaccharide-enriched fraction (CPPS) was prepared from Codonopsis pilosula root extract utilizing a procedure that entailed extraction with aqueous buffer and precipitation with ethanol. 2. After administration of CPPS in drinking water to C57BL/6 mice at a dosage of 10 mg/L for 4 weeks, the splenocytes exhibited lowered mitogenic responses to Concanavalin A (ConA) and lipopolysaccharide (LPS). The in vitro production of reactive nitrogen intermediates was inhibited. 3. However, when oral administration of CPPS was prolonged to 8 weeks, there was a potentiation of ConA-stimulated and LPS-stimulated mitogenic responses. 4. When tested under in vitro conditions, CPPS augmented the mitogenic response of splenocytes to ConA. However, there was no effect on the pinocytic activity of mouse macrophages, nor was there any proliferative activity on mouse melanoma B16 cells.

Inhibition of glycohydrolase enzymes by aqueous extracts of Chinese medicinal herbs in a microplate format

A microplate assay, for use with a variety of glycohydrolase enzymes, was developed to aid the screening of Chinese medicinal herb extracts for the presence of potential anti‐viral and anti‐lymphoma compounds. The microplate assay method described offers greater convenience, speed and reproducibility over existing methods. The enzymes tested were α‐glucosidase, β‐glucosidase and β‐glucuronidase. The assay can be easily adapted for use with other glycohydrolase enzymes. Of the 12 herb extracts examined four did not inhibit any of the enzymes (<50% inhibition), one inhibited α‐glucosidase only (>50% inhibition), six inhibited β‐glucuronidase only, and one inhibited both α‐glucosidase and β‐glucuronidase. None of the extracts were capable of inhibiting β‐glucosidase to any significant extent.

Action of α-momorcharin, a ribosome inactivating protein, on cultured tumor cell lines

Abstract 1. 1. α-Momorcharin, a glycoprotein isolated from seeds of the bitter gourd, Momordica charantia inhibited incorporation of [3H]thymidine, [3H]leucine and [3H]uridine into P388 (mouse monocyte-macrophage), J774 (Balb/c macrophage), JAR (human placental choriocarcinoma) and sarcoma