Removal of Histidine residue from ZN- a-2- glycoprotein by Carboxypeptidase enzyme using Spectrofluorimetry and Maldi-tof-Mass Spectroscopy

DOI: 10.18805/ijar.10267    | Article Id: B-420 | Page : 684-689
Citation :- Removal of Histidine residue from ZN- a-2- glycoprotein by Carboxypeptidase enzyme using Spectrofluorimetry and Maldi-tof-Mass Spectroscopy .Indian Journal Of Animal Research.2016.(50):684-689

Zain Ullah*1,2, Musa Kaleem Baloch1 and Noor Saeed Khattak3

zain_kust@yahoo.com
Address :

Department of Chemistry, Gomal University, Dera Ismail Khan, 29050, KPK, Pakistan.

Submitted Date : 22-12-2015
Accepted Date : 5-04-2016

Abstract

The present invention relates, in general, to methods developed of diagnosing and monitoring to binding with Zn-a-2-glycoprotein in order to design rational drugs. Zinc-alpha-2-glycoprotein (ZAG) is a secreted protein that is present in most bodily fluids. It is interesting because of its ability to play many important functions in the human body, including fertilization and lipid mobilization. After the discovery of this molecule, during the last 5 decades, various studies have been documented on its structure and functions but still it is considered as a protein with an unknown function. Its expression is regulated by glucocorticoids. Due to its high sequence homology with lipid-mobilizing factor and high expression in cancer cachexia, it is considered as a novel adipokine. On the other hand, its X-ray crystal structure and folding structure is similar to MHC class I antigen-presenting molecule; hence ZAG may have a role in the expression of the immune response and providing defined binding-altered mutants for cellular signaling studies and potential medical application. Spectrofluorimetry and MALDI-TOF Mass Spectroscopy showed that ZAG binds DAUDA with Kdin the micro molar range. This study will examine removal of histidine residue from recombinant ZAG protein by enzymes through Spectrofluorimetry and MALDI-TOF Mass Spectroscopy.

Keywords

Fluorescence Histidine MALDI UV absorbance Zn-a-2-glycoprotein.

References

  1. Ajayan P., Stephan O., Colliex C. and Trauth D. (1994) Aligned carbon nanotube arrays formed by cutting a polymer resin-    nanotube composite. Sceince-NewYork Then Washington. 265: 1212-1214.
  2. Arnau J., Lauritzen C., Petersen G.E. and Pedersen J. (2006) Current strategies for the use of affinity tags and tag removal for the purification of recombinant proteins. Protein Expression and Purification 48: 1-13.
  3. Balakrishnan B., Ranishree J.K., Thadikamala S. and Panchatcharam P. (2014) Purification, characterization and production optimization of a vibriocin produced by mangrove associated Vibrio parahaemolyticus. Asian Pacific Journal of Tropical Biomedicine 4: 253-61.
  4. Bing C., Bao Y., Jenkins J., Sanders P., Manieri M., Cinti S., Tisdale M.J. and Trayhurn P. (2004) Zinc-a2-glycoprotein, a lipid mobilizing factor, is expressed in adipocytes and is up-regulated in mice with cancer cachexia. Proceedings of the National Academy of Sciences of the United States of America 101: 2500-5.
  5. Brettschneider J., Mogel H., Lehmensiek V., Ahlert T., Süssmuth S., Ludolph A.C. and Tumani H. (2008) Proteome analysis of cerebrospinal fluid in amyotrophic lateral sclerosis (ALS). Neurochemical Research 33: 2358-63.
  6. Ding Z., Qu F., Guo W., Ying X., Wu M. and Zhang Y. (2007) Identification of sperm forward motility related proteins in human seminal plasma. Molecular Reproduction and Development 74: 1124-31.
  7. Gandhi N.S. and Mancera R.L. (2008) The structure of glycosaminoglycans and their interactions with proteins. Chemical Biology and Drug Design 72: 455-82.
  8. Gorelsky S.I., Basumallick L., Vura-Weis J., Sarangi R., Hodgson K.O., Hedman B., Fujisawa K. and Solomon E.I. (2005) Spectroscopic and DFT investigation of [M {HB (3, 5-i Pr2pz) 3}(SC6F5)](M= Mn, Fe, Co, Ni, Cu, and Zn) model complexes: Periodic trends in metal-thiolate bonding. Inorganic chemistry 44: 4947-60.
  9. Gundry R.L., White M.Y., Murray C.I., Kane L.A., Fu Q., Stanley B.A. and Van Eyk J.E. (2009) Preparation of Proteins and Peptides for Mass Spectrometry Analysis in a Bottom Up Proteomics Workflow. Current Protocols in Molecular Biology, 10.25. 1-10.25. 3.
  10. Hassan M.I., Waheed A., Yadav S., Singh T.P. and Ahmad F. (2008) Zinc a2-glycoprotein: a multidisciplinary protein. Molecular Cancer Research 6: 892-906.
  11. Hasson S., Al-Balushi M., Al Shaili A., Al-Busaidi J., Said E., Idris M. and Al-Jabri A. (2013) Prevalence of zinc-a 2-    glycoprotein binding peptide among Omani blood donors. Journal of Medical Laboratory 4: 21-7.
  12. Henshall S.M., Horvath L.G., Quinn D.I., Eggleton S.A., Grygiel J.J., Stricker P.D., Biankin A.V., Kench J.G. and Sutherland R.L. (2006) Zinc-alpha2-glycoprotein expression as a predictor of metastatic prostate cancer following radical prostatectomy. Journal of the National Cancer Institute 98: 1420-4.
  13. Kamoshida S., Watanabe K., SUZUKI M., Mizutani Y., Sakamoto K., Sugimoto Y., Oka T., Fukushima M. and Tsutsumi Y. (2006) Expression of cancer cachexia-related factors in human cancer xenografts: an immunohistochemical analysis. Biomedical Research 27: 275-81.
  14. Laemmli U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-5.
  15. McDermott L.C., Freel J.A., West A.P., Bjorkman P.J. and Kennedy M.W. (2006) Zn-á2-glycoprotein, an MHC class I-    related glycoprotein regulator of adipose tissues: modification or abrogation of ligand binding by site-directed mutagenesis. Biochemistry 45: 2035-41.
  16. Pal P., Rajalakshmi M., Manocha M., Sharma R., Mittal S. and Rao D. (2006) Semen quality and sperm functional parameters in fertile Indian men. Andrologia 38: 20-5.
  17. Stejskal D., Karpíšek M., Reutova H., Stejskal P., Kotolova H. and Kollár P. (2008) Determination of serum zinc-alpha-2-    glycoprotein in patients with metabolic syndrome by a new ELISA. Clinical biochemistry 41: 313-6.
  18. Vanni H., Kazeros A., Wang R., Harvey B.-G., Ferris B., De B.P., Carolan B.J., Hübner R.-H., O’Connor T.P. and Crystal R.G. (2009) Cigarette smoking induces overexpression of a fat-depleting gene AZGP1 in the human. CHEST Journal 135: 1197-208.
  19. Yee A.J., Akens M., Yang B.L., Finkelstein J., Zheng P.-S., Deng Z. and Yang B. (2007) The effect of versican G3 domain on local breast cancer invasiveness and bony metastasis. Breast Cancer Res 9: R47.

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