A. S. Kamalanathan

Bio: A. S. Kamalanathan is an academic researcher from VIT University. The author has contributed to research in topics: Affinity chromatography & Glycan. The author has an hindex of 7, co-authored 14 publications receiving 97 citations.

Molecular studies of rheumatoid factor using pseudobioaffinity membrane chromatography.

Abstract: Using pseudobioaffinity ligand L-histidine immobilized to poly(ethylene vinyl alcohol) hollow fiber membrane is an interesting approach for the purification of total IgG and its subclasses from untreated serum in a single step. Gentle adsorption and elution conditions of this chromatography system allow efficient recovery of the protein in its native form. This approach was employed for the recovery and molecular study of rheumatoid factor (RF), an anti-IgG autoantibody (AAb) that form immune complexes with autologous IgG Abs in the sera. The purity of the recovered molecule was analyzed on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE), revealed a 150-kDa IgG band and an additional ∼300 kDa band which may be RF bound IgG complex. Since RF is an AAb, the purified protein was studied for its catalytic functions like peptide, DNA, and RNA hydrolyzing activities. The substrate Pro-Phe-Arg-4-methyl-coumaryl-7-amide (PFR-MCA) hydrolyzing activity by total IgG from different patient sera was found to be greater than healthy controls. In an effort to identify the subclass specificity for the proteolytic function, the pre-purified total IgG fractions from rheumatoid arthritis (RA) sera were subjected to rechromatography using a discriminating buffer. In this experiment, the activity was found in the non-retained fractions suggesting IgG2 specificity for the catalytic function. A comparative study between different catalytic functions was performed for IgG separated from individual patient. Copyright © 2008 John Wiley & Sons, Ltd.

Proteolysis activity of IgM antibodies from rheumatoid arthritis patients' sera: evidence of atypical catalytic site.

Abstract: The IgM antibodies from rheumatoid arthritis (RA) patients' sera were screened for peptide hydrolyzing activity. Recovery of structurally intact IgM antibodies (Abs), in a single step, was achieved using a weak anion-exchange methacrylate monolith disk. The IgM Abs from patients' sera hydrolyzed the Pro-Phe-Arg-4-methyl-coumaryl-7-amide (PFR-MCA) substrate appreciably compared to the healthy donors. The apparent Km values of IgM Abs from patients' sera were between 0.4 and 0.7 mM. Furthermore, IgM Abs displayed 5 to 10-folds greater proteolysis activity than IgG Abs, recovered from the same pathological serum. The proteolysis activity, as a function, was found to be independent of IgM-RF titer value. Affinity labeling approach targeted at the catalytic site histidine was studied, using a specific irreversible inhibitor, N-α-tosyl-L-lysine chloromethyl ketone (TLCK). Despite modification of catalytic His, observation of serine protease like activity suggest presence of an atypical catalytic framework in a few pathological IgM Abs. Copyright © 2010 John Wiley & Sons, Ltd.

Overview of the Structure–Function Relationships of Mannose-Specific Lectins from Plants, Algae and Fungi

Abstract: To date, a number of mannose-binding lectins have been isolated and characterized from plants and fungi. These proteins are composed of different structural scaffold structures which harbor a single or multiple carbohydrate-binding sites involved in the specific recognition of mannose-containing glycans. Generally, the mannose-binding site consists of a small, central, carbohydrate-binding pocket responsible for the "broad sugar-binding specificity" toward a single mannose molecule, surrounded by a more extended binding area responsible for the specific recognition of larger mannose-containing N-glycan chains. Accordingly, the mannose-binding specificity of the so-called mannose-binding lectins towards complex mannose-containing N-glycans depends largely on the topography of their mannose-binding site(s). This structure⁻function relationship introduces a high degree of specificity in the apparently homogeneous group of mannose-binding lectins, with respect to the specific recognition of high-mannose and complex N-glycans. Because of the high specificity towards mannose these lectins are valuable tools for deciphering and characterizing the complex mannose-containing glycans that decorate both normal and transformed cells, e.g., the altered high-mannose N-glycans that often occur at the surface of various cancer cells.

Chemical diversity and activity profiles of HIV-1 reverse transcriptase inhibitors from plants

Abstract: Current challenges to antiretroviral therapy have opened new vistas in the search for novel drugs from natural products. This review focusses on plants as sources of inhibitors for human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. Based on a systematic search of the literature, anti-HIV-1 reverse transcriptase activity was recorded for 132 plant species in 100 genera and 51 families. Seven families comprise 52.6% of plant species with anti-reverse transcriptase activity: Lamiaceae (13.7%), Fabaceae (10.7%), Euphorbiaceae (9.9%), Clusiaceae (6.1%), Asteraceae (4.6%), Combretaceae (4.6%), and Moraceae (3.0%). The repertoire of anti-reverse transcriptase active compounds includes (-)-catechin, 1,8-cineole, 3,4-di-O-caffeoylquinic acid, 5,7-dimethoxy-6-methylflavone, apigenin, baicalein, betulinic acid, caffeic acid, cis-3-hexene-1-ol, eugenol, euscaphic acid, gallic acid, hoslunddiol, limonene, naringenin, oleanolic acid, p-cymene, pomolic acid, quinic acid, rosmarinic acid, stigmasterol, thymol, ursolic acid, α-bergamotene, α-pinene, and γ-terpinene. Among the IC50 values are 0.10 μg/ml (Uvaria angolensis), 3 μg/ml (Hemidesmus indicus), 2. 3μg/ml (Adansonia digitata), 6.24 μg/ml (Caesalpinia coriaria), 7.2 μg/ml (Terminalia sericea), 17.4 μg/ml (Hypoxis hemerocallidea), and 79 μg/ml (Moringa oleifera). The chemical diversity and activity profiles of HIV-1 reverse transcriptase inhibitors from plants reveal two recurring motifs: the structure of several active anti-reverse transcriptase compounds mimics nucleoside analogues, and numerous anti-reverse transcriptase phytochemicals have pleiotropic effects and heterogenous pharmacological benefits during infection and disease. To accelerate drug discovery and development, this review recommends the urgent need to tap into the rich vein of indigenous knowledge of putative anti-HIV/AIDS medicinal plants (reverse pharmacology), determine pan-assay interference compounds, analyze structure-activity relationships, and conduct more clinical trials.