Effect of myo-inositol(1,4,5)trisphosphate on the hydrolysis of phytic acid by phytase.

Phytase is a monomeric enzyme of molecular mass 160 kDa which catalyzes the hydrolysis of phytic acid (D-myo inositol hexakisphosphate, InsP6) in a stepwise manner to myo-inositol. The enzyme-InsPn (n = 1-6) interaction at the catalytic site has a dissociation constant in the micro molar range. There also exists in the enzyme, a non-catalytic site specific for InsP3 with dissociation constant in the nano molar range. We have probed the effect of the high affinity InsP3 binding on the dissociation constant (Kd) of the phytase-InsP6 interaction and the kinetics of hydrolysis. These studies demonstrate the effect exerted by the high affinity InsP3 binding on the catalytic site of the enzyme.

Transformation and regeneration of mung bean (Vigna radiata).

The procedure relied on a protocol in which shoot organogenesis was induced on cotyledons of mung bean genotypes selected for susceptibility to agrobacterium seems to work reproducibly if not efficiently. Approximately 4-5% of the shoots produced on the kanamycin selected cotyledons are transgenic based on assays on kanamycin resistance and GUS activity. This demonstrated that transformation and regeneration in mung bean are possible. However, raising the transformed plants in field condition is yet to be perfected.

Purification and characterization of tubulin from mung bean ( Vignaradiata).

Tubulin has been purified from mung bean seedling by Zn2+-induced polymerization. Both a- and β-subunits of mung bean tubulin are different from those of brain tubulin in electrophoretic mobility, colchicine binding and peptide map. Heterogeneity of mung bean tubulin has also been documented suggesting diversification of tubulin despite its conserved nature in general.

A single form of metallothionein is present in both heavy metal induced and neonatal chicken liver.

Multiplicity of metallothionein and their genes in higher animals are documented extensively in recent literature. In contrast, chicken liver produced apparently a single form of metallothionein upon heavy metal exposure. This protein was purified by gel filtration and ion exchange chromatography and another technique based on heat treatment and acetone fractionation, followed by ion exchange chromatography. In adult uninduced chicken liver the presence of metallothionein was below the detection limit. But, like mammalian system, chicken liver was found to contain high amount of metallothionein at neonatal stage. This naturally occurring neonatal chicken hepatic metallothionein was purified and compared with the heavy metal induced adult hepatic metallothionein. The biochemical and immunobiological comparative analysis of adult and neonatal hepatic metallothionein showed identical characteristics. The neonatal metaltothionein expressed naturally was a zinc metallothionein and unlike few other mammalian neonatal metallothionein did not contain any copper. Metallothionein was undectable in unfertilized eggs, in early embryos, and in postnatal chicken, from 4 weeks after birth. The highest level of this naturally occurring neonatal metallothionein was found in 1–4 day old neonatal liver, which was about 1·5% of the total cytosolic protein. This is the first reported evidence for the presence of ontogenically modulated expression of metallothionein in avian system. Possible biological role of neonatal metallothionein and their cellular interactions has been discussed.

Molecular biology of tubulin: Its interaction with drugs and genomic organization.

Microtubules are ubiquitous cellular structures found in eukaryotic organisms and responsible for a variety of functions. These functions include mitosis, motility, cytoskeletal architecture, intracellular transport and secretion. The major structural component of microtubules is tubulin, a dimeric protein molecule consisting of two similar but nonidentical subunits (α and β) each of about molecular weight 55,000. With the introduction of radioactive colchicine for the first time it has been reported that colchicine binds specifically to tubulin. At this point microtubule research stepped up to a new era linking microtubules with other spindle poisons which are structurally diverse as well as binding at different sites on to the tubulin heterodimer. These antimicrotubular agents have already provided valuable information regarding microtubule-mediated cellular functions and its association and dissociation phenomena. Tubulins appear to be conserved proteins based on in vitro copolymerization and comigration on polyacrylamide gel electrophoretic properties. Further, amino acid sequences of both α and β subunits from a variety of sources also appear to be mostly conserved. The evolutionary conservation of tubulin genes is highly reflected at the nucleic acid level as well. The estimation of the number of genes for tubulin and their organization in a variety of organisms have opened up a new dimension to microtubule and tubulin research. The multigene family for tubulins comprising also pseudogenes is suggestive that more than one gene for each α and β tubulin is functional in the cell. Therefore, it has been speculated that different tubulin gene products contribute to functionally different microtubules at specific stages in cell cycle and cell growth. Heterogeneity in both α and β tubulins has already been established during different stages of development of the cell. Obviously, it reflects that tubulin genes are highly regulated and this regulation might be at the transcriptional and/or translational level. Whatever is the actual control mechanism it appears that cells can detect an enhanced pool of depolymerized subunits and a rapid and specific control in tubulin gene expression at the transcriptional and/or post transcriptional level does occur.

Colchicine binding activity of rat brain polysomes.

In this communication, we report the presence of a unique colchicinebinding activity in the polysomes of rat brain. This drug-binding property, is somewhat similar to that of tubulin isolated from many sources; however, it differs in several biochemical characteristics such as (i) thermal stability of colchicine-binding site, (ii) protection of binding site by vinblastine and (iii) time required for binding equilibration. Such binding of colchicine to the polysomes is most probably due to the presence of a nascent peptide chain of tubulin in the polysome.

RNA synthesis in maturing avian erythrocyte nuclei.

The rate of RNA synthesis and its inhibition by α-amanitin in the nuclei of mature and immature avian erythrocytes are increased with the increase in ionic strength of incubation medium. Polyacrylamide gel electrophoresis indicates that heterogeneous species of RNAs are synthesised in the mature and immature erythrocyte nuclei. However, a large number of high molecular weight RNAs are synthesised in the nuclei of immature erythrocytes. Elution profiles on poly(U)- sepharose chromatography indicate that the RNAs synthesised in the nuclei of two types of cell contain poly(A) segments. Sixteen per cent of mature erythrocyte nuclear RNA syntbesised are polyadenylated, while it is 13% in immature erythrocyte nuclei. However, the total RNA synthesised is 2–3 fold higher in immature erythrocyte nuclei than that in mature erythrocyte nuclei.