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The Primary Structure of βI-Chain of Hemoglobin from Snake Sindhi Krait (Bungarus sindanus sindanus)

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Abstract

The amino acid sequence of βI-globin chain from Sindhi Krait (Bungarus sindanus sindanus) was determined to study the molecular evolution among snakes. The hemoglobin was isolated from the red blood cells and was analyzed by ion-exchange chromatography (IEX). The crude globin was subjected to reversed phased-high performance liquid chromatography (RP-HPLC) using C4 column. The N-terminal sequences of intact globin chains and tryptic peptides were determined by Edman degradation in a pulsed liquid gas phase sequencer using an online Phenylthiohydantoin analyzer. Sindhi Krait is expected to express three hemoglobin components that are composed of βII, βI, αD and αA-globin chains, as apparent by IEX, RP-HPLC and N-terminal sequence analyses. Sequence alignment and phylogenetic analyses of βI globin chain from Sindhi Krait showed closest relationship with βI globin chain from Rattlesnake, Water snake and Indigo snake. Interestingly, comparison of primary sequence of βI globin chain of Sindhi Krait with human β chain revealed 63 % similarity along with the retention of all heme contact points. Variations among the two sequences were prominent at αβ contact points and in regions directly not important for function.

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Abbreviations

IEX:

Ion-exchange

RP-HPLC:

Reversed phase-high performance liquid chromatography

Bss:

B. s. sindanus/Bungarus sindanus sindanus

TFA:

Trifluoroacetic acid

TPCK:

Tosyl-phenylalanyl-chloromethyl-ketone

PTH:

Phenylthiohydantoin

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Acknowledgments

Humera Waheed gratefully acknowledges the support from Prof. Dr. Atta-ur-Rahman (FRS) and Prof. Dr. M. Iqbal Choudhary, Director, H.E.J. Research Institute of Chemistry, ICCBS, University of Karachi, Dean Ronald P. Jordon, Professor Dr. Zahir Shaikh, College of Pharmacy, University of Rhode Island and Dr. Zafar Zaidi Trust for travel grant. Part of this research work was completed at RI-INBRE Core Facility, University of Rhode Island, supported by NIGMS, NIH Grant # 8 P20 GM103430.

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Authors and Affiliations

  1. H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan

    Humera Waheed

  2. Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, RI, 02881, USA

    Hilary Friedman & Aftab Ahmed

  3. National Center for Proteomics, University of Karachi, Karachi, 75270, Pakistan

    Syed Faraz Moin & Shamshad Zarina

  4. Chapman University School of Pharmacy, 9401 Jeronimo Road, Irvine, CA, 92618, USA

    Aftab Ahmed

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  1. Humera Waheed
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  2. Hilary Friedman
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Correspondence to Aftab Ahmed.

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Waheed, H., Friedman, H., Moin, S.F. et al. The Primary Structure of βI-Chain of Hemoglobin from Snake Sindhi Krait (Bungarus sindanus sindanus). Protein J 35, 193–201 (2016). https://doi.org/10.1007/s10930-016-9661-2

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