Comparative analysis of β-hexosaminidase and acid phosphatase from Hydra vulgaris Ind-Pune, H. vulgaris Naukuchiatal and H. magnipapillata sf-1: Localization studies of acid phosphatase and β-hexosaminidase from H. vulgaris Ind-Pune

Rohit Sai Reddy Konada; Lakshmi Surekha Krishnapati; Venugopal Ashapogu; Chung-Hung Lin; Siva Kumar Nadimpalli

doi:10.1016/j.cbpb.2019.110365

Comparative analysis of β-hexosaminidase and acid phosphatase from Hydra vulgaris Ind-Pune, H. vulgaris Naukuchiatal and H. magnipapillata sf-1: Localization studies of acid phosphatase and β-hexosaminidase from H. vulgaris Ind-Pune

Comp Biochem Physiol B Biochem Mol Biol. 2020 Jan:239:110365. doi: 10.1016/j.cbpb.2019.110365. Epub 2019 Oct 17.

Authors

Rohit Sai Reddy Konada¹, Lakshmi Surekha Krishnapati¹, Venugopal Ashapogu¹, Chung-Hung Lin², Siva Kumar Nadimpalli³

Affiliations

¹ Protein Biochemistry and Glycobiology Laboratory, Department of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad 500046, India.
² Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.
³ Protein Biochemistry and Glycobiology Laboratory, Department of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad 500046, India. Electronic address: nsksl@uohyd.ernet.in.

PMID: 31629812
DOI: 10.1016/j.cbpb.2019.110365

Abstract

The present report describes a comprehensive study on comparative biochemical characterization of two lysosomal enzymes, acid phosphatase and β-hexosaminidase in three different strains of Hydra; Hydra vulgaris Ind-Pune, H. vulgaris Naukuchiatal and H. magnipapillata sf-1 (self-feeder-1). Since morphology and habitat of Hydra effect lysosomal enzymes and their response to environmental pollutants, it would be interesting to identify them in different Hydra strains so as to use them as toxicity testing. Preliminary studies revealed a differential expression of acid phosphatase, β-hexosaminidase and β-glucuronidase in three Hydra strains. Expression of all three lysosomal enzymes in H. vulgaris Ind-Pune was low in comparison to H. vulgaris Naukuchiatal and H. magnipapillata sf-1, while their expression is comparable in H. vulgaris Naukuchiatal and H. magnipapillata sf-1. The Michaelis-Menten (K_M) values for lysosomal β-hexosaminidase using 4-nitrophenyl N-acetyl-β-D-glucosaminide as substrate were found to be 1.3 mM, 1.1 mM and 0.8 mM, respectively for H. vulgaris Ind-Pune, H. vulgaris Naukuchiatal and H. magnipapillata sf-1. For acid phosphatase using 4-nitrophenyl-phosphate as substrate, the K_M values were 0.38 mM, 1.2 mM and 0.52 mM respectively, for H. vulgaris Ind-Pune, H. vulgaris Naukuchiatal and sf-1 strains. The optimum temperature for β-hexosaminidase was 60 °C for H. vulgaris Ind-Pune, while 50 °C was observed for H. vulgaris Naukuchiatal and sf-1 strains. The optimum pH for β-hexosaminidase was found to be 6.0 for H. vulgaris Ind-Pune and H. vulgaris Naukuchiatal, and 5.0 for sf-1. The optimum temperature and pH of acid phosphatase was similar in all three strains, viz., 40 °C and 3.0, respectively. Preliminary localization studies using whole mount in situ hybridization revealed predominant endodermal expression of three enzymes in H. vulgaris Ind-Pune. Our results thus support the conservation of lysosomal hydrolases in Hydra.

Keywords: Acid phosphatase; Glucuronidase; Hexosaminidase; Hydra; Lysosomes.

Publication types

Comparative Study

MeSH terms

Acid Phosphatase / metabolism*
Animals
Hydra / enzymology*
Lysosomes / enzymology*
Species Specificity
beta-N-Acetylhexosaminidases / metabolism*

Substances

Acid Phosphatase
beta-N-Acetylhexosaminidases