Raghava Rao Jonnalagadda

Investigation on interaction of tannic acid with type I collagen and its effect on thermal, enzymatic, and conformational stability for tissue engineering applications.

Collagen is an essential component of tissues, which is the most abundant component in extracellular matrix and highly conserved across the animal kingdom. It can assemble into fiber and play an essential role in cell adhesion and growth and could be extremely useful in tissue engineering. In this study, the effect of tannic acid (TA) on the thermal, enzymatic and conformational stability of type I collagen has been investigated for the development of collagen‐based biomaterials. Interaction of TA with collagen demonstrates the role of hydrogen bonding and hydrophobic interaction in providing the thermal and enzymatic stability. Thermal analysis studies reveal that, hydrothermal stability of collagen increases as well as inhibits the breakdown of collagenase by formation of hydrogen bonds and hydrophobic interactions. TA binds to the collagen with high affinity because the structural flexibility of the collagen compensates for the structural rigidity of the phenolics. Increase in concentration of TA induces significant change in the conformation of triple helix. The free binding energy of TA with collagen‐like peptide was determined to be in the range of −9.4 to −11.2 kcal mol−1, which was calculated by using Autodock Vina software and showed numerous hydrophobic and hydrogen bond interactions. We anticipate that these collagen‐based biomaterials hold great potential for biomedical applications.

Elucidation of hydration dynamics of locust bean gum-collagen composites by impedance and thermoporometry.

The intricacy of the different parameters involved in the hydration dynamics of collagen influences its performance as biomaterials. This work presents the molecular motions of collagen originating from the solvents and locust bean gum (LBG), which reveal the changes in solvation dynamics of the biopolymers affecting the surface as well as interfacial properties. Water, as a probe liquid bound in collagen has been investigated using a combination of thermoporometry, ATR-FTIR, circular dichroic spectroscopy, dielectric spectroscopy and SEM to explore the influence of LBG on collagen with respect to static and dynamic behaviour. The relaxation process of collagen in the frequency range of 0.01 Hz to 10(5)Hz and thermoporometry results indicate that the interfacial hydration dynamics are dependent on the applied concentration of LBG. This investigation explicitly reflects the rearrangements of the structural water clusters around the charged amino acids of collagen. These results can be employed to redesign the approach towards the development of collagen based biomaterials.

Studies on the molecular significance in the interaction of bilirubin with collagen

The present investigation is aimed to understand the physiological significance of bilirubin interaction with collagen. In human skin, collagen absorbs both free bilirubin and serum bound bilirubin from the human system. Interaction between bilirubin and collagen depends on time, temperature and concentration of bilirubin. There is an increase in the aggregation rate of collagen in the presence of biliruibin. At physiological condition, 125 nM of bilirubin is the maximum concentration absorbed by per mg of collagen molecule. Bilirubin accelerates the lateral growth of collagen fibrils by shifting its rate of nucleation. Moreover, collagen-bilirubin complex exhibit a tendency to undergo adsorption onto the surface of the fibroblast cells, showing detrimental effects on fibroblasts proliferations. Based on the collagen binding assays, the binding of bilirubin to collagen is found to be electrostatic in nature, which confirms binding between the amino acid fragment of α1 (I) region of collagen and carboxyl group of bilirubin. The biotinylated bilirubin derivatives show better binding to α1 (I) chain rather than α2 (I) chains which clearly designates that bilirubin shows greater affinity to α1 chains of collagen. This novel approach directs to reduce the occurrence of bilirubin in hyperbilirubinemia patients.

Engineering D-Amino Acid Containing Collagen Like Peptide at the Cleavage Site of Clostridium histolyticum Collagenase for Its Inhibition

Collagenase is an important enzyme which plays an important role in degradation of collagen in wound healing, cancer metastasis and even in embryonic development. However, the mechanism of this degradation has not yet been completely understood. In the field of biomedical and protein engineering, the design and development of new peptide based materials is of main concern. In the present work an attempt has been made to study the effect of DAla in collagen like peptide (imino-poor region of type I collagen) on the structure and stability of peptide against enzyme hydrolysis. Effect of replacement of DAla in the collagen like peptide has been studied using circular dichroic spectroscopy (CD). Our findings suggest that, DAla substitution leads to conformational changes in the secondary structure and favours the formation of polyproline II conformation than its L-counterpart in the imino-poor region of collagen like peptides. Change in the chirality of alanine at the cleavage site of collagenase in the imino-poor region inhibits collagenolytic activity. This may find application in design of peptides and peptidomimics for enzyme-substrate interaction, specifically with reference to collagen and other extra cellular matrix proteins.

Studies on the chemico-biological characteristics of bilirubin binding with collagen.

The clinical impact of bilirubin on collagen is investigated using various physical, chemical and biological methods. Thermo gravimetric analysis and differential scanning analysis of collagen-bilirubin complex matrices indicate that crosslinking does not alter their thermal behavior of collagen. The polydispersity of collagen-bilirubin complex increases in the reacting medium suggesting that there is an increase in the number of interacting points between them. Based on the zeta potential values, the rate of mobility of interacted complex decreases by inferring the extent of binding compared to the control collagen. Emission intensity begins to increase with increase in concentration of bilirubin which ascribes the conformational changes around the aromatic amino acids in collagen. Binding is indicated by an increase in resonance units and the responses are corrected by subtraction of those obtained for native collagen. Bilirubin showed a higher affinity for collagen at a concentration of about 25 nM/mg. In this study, the association rate has been calculated which depicts the increased affinity of bilirubin to collagen. Affinity for bilirubin to collagen has been found to be 8.89×10(-3) s(-1). The greater part of binding of bilirubin to collagen is found to be electrostatic in nature. The investigation leads to comprehend the affinity of collagen-bilirubin complex during jaundice diseased tissues.

Studies on the physiological and biochemical characteristics of Salicornia brachiata: influence of saline stress due to soaking wastewater of tannery

AbstractSalicornia brachiata is an annual halophyte belonging to the Chenopodiaceae family and is distributed throughout the world. It is a green, jointed, vascular, flowering, and leafless halophytic plant that carries articulated, succulent stems. The main objectives of this study were to determine the effect of salt stress on growth characteristics, succulence, biochemical parameters, osmotic, and water relations of S. brachiata. The salt stress was induced by irrigating S. brachiata with soaking wastewater of tannery. The leather processing industry or the Tannery is known to be associated with the generation of liquid waste with high total dissolved solids (TDS), in particular, salinity. Consequently, the disposal of the tannery wastewater with high salinity has become a major concern for the tanning industry. This study evaluates the effect of tannery soaking wastewater on S. brachiata, which was grown in pots fed with soaking effluent at varying concentrations of TDS under laboratory conditions for...

Does l to d-amino acid substitution trigger helix → sheet conformations in collagen like peptides adsorbed to surfaces?

The present work reports on the structural order, self assembling behaviour and the role in adsorption to hydrophilic or hydrophobic solid surfaces of modified sequence from the triple helical peptide model of the collagenase cleavage site in type I collagen (Uniprot accession number P02452 residues from 935 to 970) using (D)Ala and (D)Ile substitutions as given in the models below: Model-1: GSOGADGPAGAOGTOGPQGIAGQRGVV GLOGQRGER. Model-2: GSOGADGP(D)AGAOGTOGPQGIAGQRGVVGLOGQRGER. Model-3: GSOGADGPAGAOGTOGPQG(D)IAGQRGVVGLOGQRGER. Collagenase is an important enzyme that plays an important role in degrading collagen in wound healing, cancer metastasis and even in embryonic development. However, the mechanism by which this degradation occurs is not completely understood. Our results show that adsorption of the peptides to the solid surfaces, specifically hydrophobic triggers a helix to beta transition with order increasing in peptide models 2 and 3. This restricts the collagenolytic behaviour of collagenase and may find application in design of peptides and peptidomimetics for enzyme-substrate interaction, specifically with reference to collagen and other extra cellular matrix proteins.