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Method for Determining Average Iron Content of Ferritin by Measuring its Optical Dispersion.

ABSTRACT: We report a method where the refractive index increments of an iron storage protein, ferritin, and apoferritin (ferritin minus iron) were measured over the wavelength range of 450-678 nm to determine the average iron content of the protein. The protein used in this study had ?3375 iron atoms per molecule. The measurement of optical dispersion over the broad wavelength range was enabled by the use of mesoporous leaky waveguides (LWs) made of chitosan. We present a facile approach for fabricating mesoporous chitosan waveguides for improving the measurement sensitivity of macromolecules such as ferritin. Mesoporous materials allow macromolecules to diffuse into the waveguide, maximizing their interaction with the optical mode and thus increasing sensitivity by a factor of ?9 in comparison to nonporous waveguides. The sensitivity was further improved and selectivity toward ferritin was achieved by the incorporation of antibodies in the waveguide. The method presented in this work is a significant advance over the state of the art method, the enzyme linked immunosorbent assay (ELISA) used in clinics, because it allows determining the average content of ferritin in a single step. The average iron content of ferritin is an important marker for conditions such as injury, inflammation, and infection. Thus, the approach presented here of measuring optical dispersion to determine the average iron content of ferritin has a significant potential to improve the point of care analysis of the protein for disease diagnosis and screening.

SUBMITTER: Gupta R

PROVIDER: S-EPMC7006959 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Method for Determining Average Iron Content of Ferritin by Measuring its Optical Dispersion.

Gupta Ruchi R Alamrani Nasser A NA Greenway Gillian M GM Pamme Nicole N Goddard Nicholas J NJ

Analytical chemistry 20190514 11

We report a method where the refractive index increments of an iron storage protein, ferritin, and apoferritin (ferritin minus iron) were measured over the wavelength range of 450-678 nm to determine the average iron content of the protein. The protein used in this study had ∼3375 iron atoms per molecule. The measurement of optical dispersion over the broad wavelength range was enabled by the use of mesoporous leaky waveguides (LWs) made of chitosan. We present a facile approach for fabricating ...[more]

PMID: 31059232

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Project description:Iron is an indispensable element for plant growth and defense and hence it is essential to improve the plant's ability to accumulate iron. Besides, it is also an important aspect for human health. In view of this, we attempted to increase the iron content in banana cultivar Rasthali using MusaFer1 as a candidate gene. Initially, the expression of all five genes of the MusaFer family (MusaFer1-5) was quantified under iron-excess and -deficient conditions. The supplementation of 250 and 350 ?M iron enhanced expression of all MusaFer genes; however, MusaFer1 was increased maximally by 2- and 4- fold in leaves and roots respectively. Under iron deficient condition, all five MusaFer genes were downregulated, indicating their iron dependent regulation. In MusaFer1 overexpressing lines, iron content was increased by 2- and 3-fold in leaves and roots respectively, as compared with that of untransformed lines. The increased iron was mainly localized in the epidermal regions of petiole. The analysis of MusaFer1 promoter indicated that it might control the expression of iron metabolism related genes and also other genes of MusaFer family. MusaFer1 overexpression led to downregulated expression of MusaFer3, MusaFer4 and MusaFer5 in transgenic leaves which might be associated with the plant's compensatory mechanism in response to iron flux. Other iron metabolism genes like Ferric reductase (FRO), transporters (IRT, VIT and YSL) and chelators (NAS, DMAS and NAAT) were also differentially expressed in transgenic leaf and root, suggesting the multifaceted impact of MusaFer1 towards iron uptake and organ distribution. Additionally, MusaFer1 overexpression increased plant tolerance against methyl viologen and excess iron which was quantified in terms of photosynthetic efficiency and malondialdehyde content. Thus, the study not only broadens our understanding about iron metabolism but also highlights MusaFer1 as a suitable candidate gene for iron fortification in banana.

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Method for Determining Average Iron Content of Ferritin by Measuring its Optical Dispersion.

Publications

Method for Determining Average Iron Content of Ferritin by Measuring its Optical Dispersion.

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