The presence of sialoglycoproteins (SGps) or glycophorins in the membranes from goat (Capra aegagrus hircus), buffalo (Bubalus bubalis bubalis) and pig (Sus scrofa domestica) erythrocytes in comparison with human (Homo sapiens) erythrocytes was investigated by partial purification and further analysis by Sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-pAGE) followed by sialoglycoprotein specific pAS stain. The results show that mammalian erythrocytes possess prominent differences in the SGps numbers and molecular weights. The solubility behaviour of goat, buffalo, pig and cow erythrocyte membrane proteins and glycophorins was studied by solubilization of membranes with non-ionic detergent (Triton – X 100). The extraction of erythrocyte membranes with Triton X-100 followed by SDS-pAGE, showed the differences in the extent of solubilization of integral proteins including glycophorins.

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Action of proteinases viz. trypsin and chymotrypsin, and neuraminidase on intact erythrocyte membrane proteins and glycophorins (sialoglycoproteins) exposed to cell surface and its impact on lectin (concanavalin A)-mediated agglutination were studied in Homo sapiens (human), Capra aegagrus hircus (goat) and Bubalus bubalis (buffalo). proteinase action(s) on human and buffalo erythrocyte surface membrane proteins and glycophorins showed similarity but was found different in goat. Significant differences in erythrocyte agglutinability with concanavalin A can be attributed to differences in membrane composition and alterations in the surface proteins after enzyme treatment. A direct correlation was found between degradation of glycophorins and concanavalin A agglutinability. Action of neuraminidase specifically indicated the negative role of cell surface sialic acids in determining concanavalin A agglutinability of goat and buffalo erythrocytes, similar to human.

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The influence of thermal stress on the association between human erythrocyte membranes and cytosolic proteins was studied by exposing erythrocyte suspensions and whole blood to different elevated temperatures. Membranes and cytosolic proteins obtained from unheated and heat stressed erythrocytes were analyzed by electrophoresis, followed by mass spectrometric identification. The heat-induced association of cytosolic proteins was restricted to the Triton shells (membrane skeleton/cytoskeleton). Similar results were observed when Triton shells derived from unheated erythrocyte membranes were incubated with an unheated erythrocyte cytosolic fraction at elevated temperatures.

Four M.Phil and Sixteen M.Sc. students were supervised for dissertation work.

List of Publications :-

1. The Red Cell Membrane Structure : Some features of interest to Hematologists. N.G.Mehta, S.M.Gokhale and R.D.Kalraiya Indian J. Hematol. Vol. 3, 41-48, (1985). ISSN – 0971-4502
2. Glycophorin A interferes in the agglutination of human erythrocytes by concanavalin A. Explanation for the requirement of enzymatic predigestion. S.M.Gokhale and N.G.Mehta Biochem. J.(U.K.), 241, 505-511, (1987). ISSN – 0264-6021
3. Concanavalin A-agglutinability of membrane-skeleton-free vesicles and aged cellular remnants derived from human erythrocytes. Is the membrane skeleton required for aggiutination ? S.M.Gokhale and N.G.Mehta Biochem. J.(U.K.), 241, 513-520, (1987). ISSN – 0264-6021
4. Concanavalin A binding to human erythrocytes leads to alterations in properties of the membrane skeleton. S.M.Gokhale and N.G.Mehta Biochem. J.(U.K.), 241, 521-525, (1987). ISSN – 0264-6021
5. Endogenous factors determining the agglutinability of erythrocytes with concanavalin A. N.G.Mehta, S.M.Gokhale and K.N.Pestonjamasp Indian J. Biochem. Biophys. 25, 14-19, (1988). ISSN – 0301-1208
6. Membrane skeleton and transbilayer phospholipid organization in heat-treated human erythrocytes. S.R.P.Gudi, A.Kumar, V.Bhakuni, S.M.Gokhale and C.M.Gupta Biochim. Biophys. Acta 1023, 63-72, (1990). ISSN – 0005-2736
7. Heat-induced alterations in monkey erythrocyte membrane phospholipid organization and skeletal protein structure and interactions. A.Kumar, S.R.P.Gudi, S.M.Gokhale, V.Bhakuni and C.M.Gupta Biochim. Biophys. Acta 1030, 269-278, (1990). ISSN – 0005-2736
8. The role of the membrane skeleton in concanavalin A-mediated agglutination of human erythrocytes. K.N. Pestonjamasp, S.M. Gokhale and N.G.Mehta Biotechnol Appl Biochem.12 (5), 544-549, (1990) ISSN – 0885-4513
9. Solubility behavior of integral proteins and glycophorins of mammalian erythrocyte membrane. Savita Sharma and S. M. Gokhale Asian J. Exp. Biol. Sci. Vol. 2 (3), 449-454 (2011) ISSN - 0975-5845
10. Sialoglycoproteins of mammalian erythrocyte membranes: A comparative study. Savita Sharma and Sadashiv M. Gokhale Asian-Australian J. Anim. Sci. Vol. 24 (12), 1666-1673 (2011) Online ISSN - 1976-5517 Print ISSN - 1011-2367
11. Differential actions of proteinases and neuraminidase on mammalian erythrocyte surface and its impact on erythrocyte agglutination by concanavalin A. Gen. Physiol. Biophys. 31, 457–468 (2012) ISSN - 0231-5882
12. Identification of human erythrocyte cytosolic proteins associated with plasma membrane during thermal stress. Savita Sharma, Surekha M. Zingde and Sadashiv M. Gokhale J. Membrane Biology 246 (8), 591-607 (2013) Online ISSN- 1432-1424 Print ISSN- 0022-2631
13. Marker assisted backcross selection for genetic removal of lipoxygenase-2 from popular soybean (Glycine max) variety JS 97-52: Parental polymorphism survey and hybridity validation. Reena Rawal, Vineet Kumar, Anita Rani, S M Gokhale and S M Husain Indian Journal of Agricultural Sciences 84 (3): 414–7, March 2014 ISSN: 0019-5022 Impact factor- 0.141 (2014), 0.231 (2017/18)
14. A comparative study of the membrane proteins of some mammalian erythrocytes identified by mass spectrometry. Savita Sharma, Vinny Punjabi, Surekha M. Zingde and Sadashiv M. Gokhale J. Membrane Biology 247(11),1181–1189, Oct 2014 Online ISSN: 1432-1424 Print ISSN: 0022-2631 (Springer) Impact factor- 2.457 (2014), 1.638 (2017/18)
15. Enzymatic cleavage of cell surface proteins of pig and cow erythrocytes and its effect on concanavalin-mediated agglutinability. Savita Sharma and Sadashiv M. Gokhale Indian J Biochem Biophys 51(5) 378-387,Oct 2014 Online ISSN: 0975-0959 Print ISSN: 0301-1208 Impact factor- 0.871 (2014), 0.385 (2017/18)
16. Alkyloxy carbonyl modified hexapeptides as a high affinity compounds for Wnt5A protein in the treatment of psoriasis. Ankit Kelotra, Sadashiv M Gokhale, Seema Kelotra, Vaidehi Mukadam, Komal Nagwanshi, Srinivas Bandaru, Anuraj Nayarisseri & Anil Bidwai Bioinformation 10(12): 739-745, Dec 2014 Online ISSN: 0973-2063 Print ISSN: 0973-8894
17. Morphological trait variations in the west Himalayan (India) populations of Arabidopsis thaliana along altitudinal gradients. Akanksha Singh, Antariksh Tyagi, Abhinandan Mani Tripathi, Sadashiv M. Gokhale, Nandita Singh and Sribash Roy. Current Science 108 (12): 2213-2222, June 2015 Online ISSN: 0011-3891 Impact factor- 0.967 (2015), 0.883 (2017/18)
18. Effect of free radical generating system- Adenosine Triphosphate/Ferrous Sulphate/Hydrogen Peroxide on human erythrocytes. Vinny Punjabi and Sadashiv M Gokhale Inter. J. of Contemporary Res. and Rev. 8 (7), 20184-20192, July 2017 Online ISSN: 0976 – 4852
19. Structure of Asp-bound peptidase E from Salmonella enterica: Active site at dimer interface illuminates Asp recognition. Pooja Yadav, Venuka Durani Goyal, Neeraj K. Gaur, Ashwani Kumar, Sadashiv M. Gokhale and Ravindra D. Makde FEBS Letters- doi:10.1002/1873-3468.13247, Sept 2018 Online ISSN: 1873-3468 Print ISSN: 0014-5793 Impact factor: 2.999 (2017/18)