Crocodilian τ-Crystallin: Overexpression, Purification, and Characterization

Mishra, Anurag; Chandrashekhar, Reena; Aggarwal, Ramesh K.; Sharma, Yogendra

doi:10.1006/prep.2001.1609

Cited by 5 publications

(6 citation statements)

References 12 publications

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“…Although τ‐crystallin has been identified in the adult bovine cornea, its expression during embryonic development has not been studied. We have followed its expression in crocodile cornea at various stages of embryonic development by western blot analysis and immunofluorescence using a polyclonal rat antiserum raised against recombinant τ‐crystallin cloned from a crocodile lens [29]. The western blot data showed that τ‐crystallin is present, albeit at a lower concentration, in the corneal extract of the cornea from the stage 21 embryo and its expression increased during further stages of development as seen from the intensity of the band (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Purification of recombinant s-crystallin and generation of polyclonal s-crystallin/a-enolase and TIM antibodies Recombinant s-crystallin was cloned and overexpressed from crocodilian lens as described earlier [29]. Antibody against recombinant a-enolase ⁄ s-crystallin was raised in a six-month-old mouse using the standard procedure.…”

Section: Protein Identificationsmentioning

confidence: 99%

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Triose phosphate isomerase, a novel enzyme‐crystallin, and τ‐crystallin in crocodile cornea

Kathiresan¹,

Krishnan²,

Krishnakumar

et al. 2006

The FEBS Journal

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Crystallins are defined as lens structural proteins, which are classified as ubiquitous a-, b-and c-crystallins, and taxon-specific crystallins [1]. The transparency and refractive properties of lens depends on crystallins. Taxon-specific crystallins are generally enzymes recruited as structural proteins in the lens to perform specialized functions, i.e., maintaining lens transparency [2]. Some examples of the species-specific crystallins are d-and e-crystallins present in birds and reptiles, s-crystallins in turtle and reptiles, q-crystallins in frog and camel lenses [3]. Several of these crystallins are metabolic enzymes: d-crystallin is argininosuccinate lyase [4]; s-crystallin is a-enolase [5]. They are believed to be recruited by gene sharing or by gene duplication [2,6].Some enzymes such as aldehyde dehydrogenase class 3 (ALDH3), aldehyde dehydrogenase class 1 Several enzymes are known to accumulate in the cornea in unusually high concentrations. Based on the analogy with lens crystallins, these enzymes are called corneal crystallins, which are diverse and species-specific. Examining crystallins in lens and cornea in multiple species provides great insight into their evolution. We report data on major proteins present in the crocodile cornea, an evolutionarily distant taxon. We demonstrate that s-crystallin ⁄ a-enolase and triose phosphate isomerase (TIM) are among the major proteins expressed in the crocodile cornea as resolved by 2D gel electrophoresis and identified by MALDI-TOF. These proteins might be classified as putative corneal crystallins. s-Crystallin, known to be present in turtle and crocodile lens, has earlier been identified in chicken and bovine cornea, whereas TIM has not been identified in the cornea of any species. Immunostaining showed that s-crystallin and TIM are concentrated largely in the corneal epithelium. Using western blot, immunofluorescence and enzymatic activity, we demonstrate that high accumulation of s-crystallin and TIM starts in the late embryonic development (after the 24th stage of embryonic development) with maximum expression in a two-week posthatched animal. The crocodile corneal extract exhibits significant a-enolase and TIM activities, which increases in the corneal extract with development. Our results establishing the presence of s-crystallin in crocodile, in conjunction with similar reports for other species, suggest that it is a widely prevalent corneal crystallin. Identification of TIM in the crocodile cornea reported here adds to the growing list of corneal crystallins.Abbreviations ALDH, aldehyde dehydrogenase; IEF, isoelectric focusing; TIM, triose phosphate isomerase.

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Section: Resultsmentioning

confidence: 99%

Section: Protein Identificationsmentioning

confidence: 99%

Triose phosphate isomerase, a novel enzyme‐crystallin, and τ‐crystallin in crocodile cornea

Kathiresan¹,

Krishnan²,

Krishnakumar

et al. 2006

The FEBS Journal

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“…squid and scallop) have high concentrations of glutathione S-transferase and aldehyde dehydrogenase in their corneas that are also lens crystallins (21,22). -Crystallin/␣-enolase is a known example of a lens and the corneal crystallin in some species (6,14,40,41). The accumulation of ubiquitous lens ␣-, ␤-, and ␥-crystallins in a cornea has never been reported.…”

Section: ␣B- ␤B2- and ␥Ii-crystallins Localize In Corneal Epithelialmentioning

confidence: 99%

Ubiquitous Lens α-, β-, and γ-Crystallins Accumulate in Anuran Cornea as Corneal Crystallins

Krishnan¹,

Kathiresan²,

Raman³

et al. 2007

Journal of Biological Chemistry

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Corneal epithelium is known to have high levels of some metabolic enzymes such as aldehyde dehydrogenase in mammals, gelsolin in zebrafish, and ␣-enolase in several species. Analogous to lens crystallins, these enzymes and proteins are referred to as corneal crystallins, although their precise function is not established in any species. Although it is known that after lentectomy, the outer cornea undergoes transdifferentiation to regenerate a lens only in anuran amphibians, major proteins expressed in an anuran cornea have not been identified. This study therefore aimed to identify the major corneal proteins in the Indian toad (Bufo melanostictus) and the Indian frog (Rana tigrina). Soluble proteins of toad and frog corneas were resolved on two-dimensional gels and identified by matrix-assisted laser desorption ionization time-of-flight/time-of-flight and electrospray ionization quadrupole time-of-flight. We report that anuran cornea is made up of the full complement of ubiquitous lens ␣-, ␤-, and ␥-crystallins, mainly localized in the corneal epithelium. In addition, some taxon-specific lens crystallins and novel proteins, such as ␣-or ␤-enolase/-crystallin, were also identified. Our data present a unique case of the anuran cornea where the same crystallins are used in the lens and in the cornea, thus supporting the earlier idea that crystallins are essential for the visual functions of the cornea as they perform for the lens. High levels of lens ␣-, ␤-, and ␥-crystallins have not been reported in the cornea of any species studied so far and may offer a possible explanation for their inability to regenerate a lens after lentectomy. Our data that anuran cornea has an abundant quantity of almost all the lens crystallins are consistent with its ability to form a lens, and this connection is worthy of further studies.

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“…, tau-crystallin (spot 74) and α-enolase (spots 59, 77 and 78), have also been found in amphibians and reptiles. Tau-crystallin is a taxon-restricted crystallin found in the eye lenses of reptiles and a few avian species but is presumably absent in mammals (Mishra et al , 2002). The level of tau-crystallin expression in the lens varies among species, e.g.…”

Section: Discussionmentioning

confidence: 99%

“…The level of tau-crystallin expression in the lens varies among species, e.g. , in crocodiles it is the least abundant crystallin and is present in trace amounts (Williams et al , 1985; Mishra et al , 2002). Interestingly, tau-crystallin had a relatively high expression in E. taeniura liver.…”

Section: Discussionmentioning

confidence: 99%

Proteomic profiling of liver from Elaphe taeniura, a common snake in eastern and southeastern Asia

et al. 2013

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Snake liver has been implicated in the adaptation of snakes to a variety of habitats. However, to date, there has been no systematic analysis of snake liver proteins. In this study, we undertook a proteomic analysis of liver from the colubrid snake Elaphe taeniura using a combination of two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flightmass spectrometry (MALDI-TOF MS). We also constructed a local protein sequence database based on transcriptome sequencing to facilitate protein identification. Of the 268 protein spots revealed by 2-DE 109 gave positive MS signals, 84 of which were identified by searching the NCBInr, Swiss-Prot and local databases. The other 25 protein spots could not be identified, possibly because their transcripts were not be stable enough to be detected by transcriptome sequencing. GO analysis showed that most proteins may be involved in binding, catalysis, cellular processes and metabolic processes. Forty-two of the liver proteins identified were found in other reptiles and in amphibians. The findings of this study provide a good reference map of snake liver proteins that will be useful in molecular investigations of snake physiology and adaptation.

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Crocodilian τ-Crystallin: Overexpression, Purification, and Characterization

Cited by 5 publications

References 12 publications

Triose phosphate isomerase, a novel enzyme‐crystallin, and τ‐crystallin in crocodile cornea

Triose phosphate isomerase, a novel enzyme‐crystallin, and τ‐crystallin in crocodile cornea

Ubiquitous Lens α-, β-, and γ-Crystallins Accumulate in Anuran Cornea as Corneal Crystallins

Proteomic profiling of liver from Elaphe taeniura, a common snake in eastern and southeastern Asia

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