Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Spectroscopic Studies on the Conformational Stability of Hemocyanin of Pila virens (Lamarck, 1822) in the Presence of Temperature and Detergents

Spectroscopic Studies on the Conformational Stability of Hemocyanin of Pila virens (Lamarck,... Gastropods are endowed with a soluble copper containing, blue coloured respiratory protein known as hemocyanin. In the present study, for the first time, hemocyanin from Pila virens, a fresh water gastropod endemic to Indian sub-continent was studied for its conformational stability in the presence of temperature and various detergents by different spectroscopic techniques viz. UV–Vis, fluorescence and circular dichroism spectroscopy. Results from UV–Vis spectroscopy revealed that, absorbance corresponding to active site moieties were decreased with increase in temperature from 30 to 70 °C. Whereas ‘thermal reversibility’ studies showed the increase in absorbance of active site with decrease in temperature from 70 to 20 °C (is an indicative of the characteristic feature of hemocyanins—that oxygen is reversibly bound to the copper in the active site). Results from CD spectroscopy revealed that the temperature-induced conformational changes on the secondary structure of hemocyanin was reversible up to 70 °C. Beyond 80 °C, the protein was completely denatured and assumed an increased random coil conformation. Hemocyanin in the presence of high concentration of detergents lost its structure as evident from UV–Vis fluorescence (evident from the red shifts in the emission maxima accompanied by quenching in emission intensities) and CD spectroscopy. Stability of active site moieties in the presence of different detergents was also studied by UV–Vis spectroscopy. The biophysical techniques gave insights into the varied effects of each of the different denaturants on the conformational stability of hemocyanin from Pila virens. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Proceedings of the Zoological Society Springer Journals

Spectroscopic Studies on the Conformational Stability of Hemocyanin of Pila virens (Lamarck, 1822) in the Presence of Temperature and Detergents

Download PDF
10 pages

Loading next page...
 
/lp/springer-journals/spectroscopic-studies-on-the-conformational-stability-of-hemocyanin-of-Vg50T0yjtQ
Publisher
Springer Journals
Copyright
Copyright © Zoological Society, Kolkata, India 2019
Subject
Life Sciences; Life Sciences, general; Zoology; Animal Anatomy / Morphology / Histology; Animal Genetics and Genomics; Biodiversity; Conservation Biology/Ecology
ISSN
0373-5893
eISSN
0974-6919
DOI
10.1007/s12595-019-00317-2
Publisher site
See Article on Publisher Site

Abstract

Gastropods are endowed with a soluble copper containing, blue coloured respiratory protein known as hemocyanin. In the present study, for the first time, hemocyanin from Pila virens, a fresh water gastropod endemic to Indian sub-continent was studied for its conformational stability in the presence of temperature and various detergents by different spectroscopic techniques viz. UV–Vis, fluorescence and circular dichroism spectroscopy. Results from UV–Vis spectroscopy revealed that, absorbance corresponding to active site moieties were decreased with increase in temperature from 30 to 70 °C. Whereas ‘thermal reversibility’ studies showed the increase in absorbance of active site with decrease in temperature from 70 to 20 °C (is an indicative of the characteristic feature of hemocyanins—that oxygen is reversibly bound to the copper in the active site). Results from CD spectroscopy revealed that the temperature-induced conformational changes on the secondary structure of hemocyanin was reversible up to 70 °C. Beyond 80 °C, the protein was completely denatured and assumed an increased random coil conformation. Hemocyanin in the presence of high concentration of detergents lost its structure as evident from UV–Vis fluorescence (evident from the red shifts in the emission maxima accompanied by quenching in emission intensities) and CD spectroscopy. Stability of active site moieties in the presence of different detergents was also studied by UV–Vis spectroscopy. The biophysical techniques gave insights into the varied effects of each of the different denaturants on the conformational stability of hemocyanin from Pila virens.

Journal

Proceedings of the Zoological Society – Springer Journals

Published: Sep 5, 2020

References

  • A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
    Bradford, MM
  • Diverse immune functions of hemocyanins
    Coates, CJ; Nairn, J
  • Antifungal peptides are generated from the C terminus of Shrimp hemocyanin in response to microbial challenge
    Destoumieux-Garzon, D; Saulnier, D; Garnier, J; Jouffrey, C; Bulet, P; Bachere, E
  • Structural and conformational stability of hemocyanin from the garden snail Cornu aspersum
    Dolashki, A; Velkova, L; Voelter, W; Dolashka, P
  • Guanidinium chloride induced unfolding of a hemocyanin subunit from Carcinus aestuarii I. Apo form
    Favilla, R; Goldoni, M; Mazzini, A; Muro, PD; Salvato, B; Beltramini, M
  • Guanidinium chloride induced unfolding of a hemocyanin subunit from Carcinus aestuarii II. Holo form
    Favilla, R; Goldoni, M; Signore, FD; Muro, PD; Salvato, B; Beltramini, M
  • Conformational states of the Rapana thomasiana hemocyanin and its substructures studied by dynamic light scattering and time-resolved fluorescence spectroscopy
    Georgieva, D; Schwark, D; Nikolov, P; Idakieva, K; Parvanova, K; Dierks, K; Genov, N; Betzel, C
  • Hemocyanin with phenoloxidase activity in the chitin matrix of the crayfish gastrolith
    Glazer, L; Tom, M; Weil, S; Roth, Z; Khalaila, I; Mittelman, B; Sagi, A
  • Irreversible thermal denaturation of hemocyanin of Helix pomatia and its substructures studied by differential scanning calorimetry
    Idakieva, K; Gielens, C; Siddiqui, NI; Doumanova, L; Vaseva, B; Kostov, G; Shnyrov, VL
  • Reversible heat inactivation of copper sites precedes thermal unfolding of molluscan (Rapana thomasiana) hemocyanin
    Idakieva, K; Meersman, F; Gielens, C
  • Spectroscopic properties and conformational stability of Concholepas concholepas hemocyanin
    Idakieva, K; Nikolov, P; Chakarska, I; Genov, N; Shnyrov, VL
  • Differential scanning calorimetry of the irreversible denaturation of Rapana thomasiana (marine snail, Gastropod) hemocyanin
    Idakieva, K; Parvanova, K; Todinova, S
  • Fluorescence properties and conformational stability of β-hemocyanin of Helix pomatia
    Idakieva, K; Siddiqui, NI; Parvanova, K; Nikolov, P; Gielens, C
  • Effects of guanidine hydrochloride on the conformation and enzyme activity of streptomycin adenylyltransferase monitored by circular dichroism and fluorescence spectroscopy
    Jana, S; Chaudhuri, TK; Deb, JK
  • Moult cycle specific differential gene expression profiling of the crab Portunus pelagicus
    Kuballa, AV; Holton, TA; Paterson, B; Elizur, A
  • Processing of an antibacterial peptide from hemocyanin of the freshwater crayfish Pacifastacus leniusculus
    Lee, SY; Lee, BL; Soderhall, K
  • Thermal denaturation and protein stability analysis of Haliotis rubra hemocyanin
    Marshall, G; Valtchev, P; Dehghani, F; Gomes, VG
  • Purification, spectroscopic characterization and o-diphenoloxidase activity of hemocyanin from a freshwater gastropod: Pila globosa
    Naresh, KN; Krupanidhi, S; Rajan, SS
  • Structural insights into the interaction between molluscan hemocyanins and phenolic substrates: An in silico study using docking and molecular dynamics
    Naresh, KN; Sreekumar, A; Rajan, SS
  • C-terminal functional unit of Rapana thomasiana (marine snail, gastropod) hemocyanin isoform RtH1: Isolation and characterization
    Parvanova, K; Idakieva, K; Todinova, S; Genov, N
  • The physiological significance of respiratory proteins in invertebrates
    Paul, R; Pirow, R
  • Phenoloxidase activity of Helix aspersa maxima (Garden snail, gastropod) hemocyanin
    Raynova, Y; Doumanova, L; Idakieva, KN
  • SDS-induced phenoloxidase activity of Helix aspersa maxima hemocyanin
    Raynova, Y; Todinova, S; Yordanov, D; Idakieva, K
  • Denaturation studies by fluorescence and quenching of thermophilic protein NAD-glutamate dehydrogenase from Thermus thermophilus HB8
    Ruiz, JL; Ferrer, J; Pire, C; Llorca, FI; Bonete, MJ
  • Effect of denaturants on the structure and activity of 3-hydroxybenzoate-6-hydroxylase
    Sumathi, S; Dasgupta, D
  • Keyhole limpet hemocyanin: Structural and functional characterization of two different subunits and multimers
    Swerdlow, RD; Ebert, RF; Lee, P; Bonaventura, C; Miller, KI
  • Irreversible thermal denaturation of Helix aspersa maxima hemocyanin
    Todinova, S; Raynova, Y; Idakieva, K
  • Calorimetric study of Helix aspersa Maxima hemocyanin isoforms
    Todinova, S; Raynova, Y; Idakieva, K
  • Acid-induced unfolding of didecameric keyhole limpet hemocyanin: Detection and characterizations of decameric and tetrameric intermediate states
    Varshney, A; Ahmad, B; Rabbani, G; Kumar, V; Yadav, S; Khan, RH
  • Rapana venosa hemocyanin with antiviral activity
    Velkova, L; Todorov, D; Dimitrov, I; Shishkov, S; Beeumen, JV; Dolashka-Angelova, P
  • Thermal stability of homologous functional units of Helix pomatia hemocyanin does not correlate with carbohydrate content
    Yesilyurt, B; Gielens, C; Meersman, F