Extraction, purification and characterization of the crystallin protein of cataractous eye lens nucleus

Muhammad Sher, Ayesha Hameed, Sobia Noreen, Muhammad Fayyaz-Ur-Rehman, Muhammad Ajaz Hussain, Bukhari Syed Nasir Abbas

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The purpose of this study is to separate and identify the crystallin protein present in the nucleus of a human cataractous eye lens. Cataractous lenses were collected from different eye hospitals from patients of different etiologies with ages between 40 and 80 years. Lens nucleus proteins were extracted into four fractions on the basis of their solubility in different media by applying a reported method. These fractions were buffer-soluble proteins (PS), urea-soluble proteins (PU), yellow fraction proteins (PY) and insoluble proteins (PI). All three soluble fractions were subjected to HPLC and GPC analysis. Both HPLC and GPC analysis showed that each fraction contains α-, β- and γ-crystallins, a major class of protein present in the lenses of vertebrates. Various chromatographic parameters including precision, accuracy and linearity have been evaluated. Studies of water-insoluble crystallins using sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) have demonstrated extreme homogeneity with evidence of major components with molecular masses of 18-70 kDa, similar to the crystallin of the water-soluble portion. The method was found to be suitable for the analysis of various isomers of crystallin protein present in human cataractous eye lens nuclei. The detailed results of the GPC are discussed. This study provides the first HPLC and GPC analysis of a human cataractous eye lens nucleus.

Original languageEnglish
Pages (from-to)6392-6397
Number of pages6
JournalThe Analyst
Volume140
Issue number18
DOIs
Publication statusPublished - 21 Sep 2015

Fingerprint

Crystallins
Purification
purification
Lenses
Proteins
protein
Crystalline Lens
High Pressure Liquid Chromatography
Water
etiology
Molecular mass
Solubility
linearity
Polyacrylates
Electrophoresis
Vertebrates
Urea
Polyacrylamide Gel Electrophoresis
Isomers
Buffers

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Electrochemistry
  • Biochemistry
  • Environmental Chemistry

Cite this

Sher, M., Hameed, A., Noreen, S., Fayyaz-Ur-Rehman, M., Hussain, M. A., & Syed Nasir Abbas, B. (2015). Extraction, purification and characterization of the crystallin protein of cataractous eye lens nucleus. The Analyst, 140(18), 6392-6397. https://doi.org/10.1039/c5an01212k

Extraction, purification and characterization of the crystallin protein of cataractous eye lens nucleus. / Sher, Muhammad; Hameed, Ayesha; Noreen, Sobia; Fayyaz-Ur-Rehman, Muhammad; Hussain, Muhammad Ajaz; Syed Nasir Abbas, Bukhari.

In: The Analyst, Vol. 140, No. 18, 21.09.2015, p. 6392-6397.

Research output: Contribution to journalArticle

Sher, M, Hameed, A, Noreen, S, Fayyaz-Ur-Rehman, M, Hussain, MA & Syed Nasir Abbas, B 2015, 'Extraction, purification and characterization of the crystallin protein of cataractous eye lens nucleus', The Analyst, vol. 140, no. 18, pp. 6392-6397. https://doi.org/10.1039/c5an01212k
Sher M, Hameed A, Noreen S, Fayyaz-Ur-Rehman M, Hussain MA, Syed Nasir Abbas B. Extraction, purification and characterization of the crystallin protein of cataractous eye lens nucleus. The Analyst. 2015 Sep 21;140(18):6392-6397. https://doi.org/10.1039/c5an01212k
Sher, Muhammad ; Hameed, Ayesha ; Noreen, Sobia ; Fayyaz-Ur-Rehman, Muhammad ; Hussain, Muhammad Ajaz ; Syed Nasir Abbas, Bukhari. / Extraction, purification and characterization of the crystallin protein of cataractous eye lens nucleus. In: The Analyst. 2015 ; Vol. 140, No. 18. pp. 6392-6397.
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