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Biophysical characterization and molecular phylogeny of human KIN protein

  • José Renato Pattaro Júnior
  • Ícaro Putinhon Caruso
  • Quirino Alves de Lima Neto
  • Francisco Ferreira Duarte Junior
  • Fabiana dos Santos Rando
  • Edileusa Cristina Marques Gerhardt
  • Maria Aparecida Fernandez
  • Flávio Augusto Vicente SeixasEmail author
Original Article
  • 69 Downloads

Abstract

The DNA/RNA-binding KIN protein was discovered in 1989, and since then, it has been found to participate in several processes, e.g., as a transcription factor in bacteria, yeasts, and plants, in immunoglobulin isotype switching, and in the repair and resolution of double-strand breaks caused by ionizing radiation. However, the complete three-dimensional structure and biophysical properties of KIN remain important information for clarifying its function and to help elucidate mechanisms associated with it not yet completely understood. The present study provides data on phylogenetic analyses of the different domains, as well as a biophysical characterization of the human KIN protein (HSAKIN) using bioinformatics techniques, circular dichroism spectroscopy, and differential scanning calorimetry to estimate the composition of secondary structure elements; further studies were performed to determine the biophysical parameters ΔHm and Tm. The phylogenetic analysis indicated that the zinc-finger and winged helix domains are highly conserved in KIN, with mean identity of 90.37% and 65.36%, respectively. The KOW motif was conserved only among the higher eukaryotes, indicating that this motif emerged later on the evolutionary timescale. HSAKIN has more than 50% of its secondary structure composed by random coil and β-turns. The highest values of ΔHm and Tm were found at pH 7.4 suggesting a stable structure at physiological conditions. The characteristics found for HSAKIN are primarily due to its relatively low composition of α-helices and β-strands, making up less than half of the protein structure.

Keywords

KIN (Kin17) protein Phylogeny Circular dichroism DSC analysis Tumor marker 

Notes

Acknowledgements

This work was supported by Fundação Araucária (Grant Numbers 147/14 and 40/16), Coordination for the Improvement of Higher Education Personnel—Brazil (CAPES, code 001), and the National Council for Scientific and Technological Development—Brazil (CNPq Grant Number 305960/2015-6). Dr. Fátima Pereira de Souza and Dr. Marcelo Andrés Fossey are gratefully acknowledged for the helpful discussion, and the authors thank IBILCE/UNESP and UFPR for the use of facilities.

Supplementary material

249_2019_1390_MOESM1_ESM.docx (5.9 mb)
Supplementary file1 (DOCX 6006 kb)

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Copyright information

© European Biophysical Societies' Association 2019

Authors and Affiliations

  • José Renato Pattaro Júnior
    • 1
  • Ícaro Putinhon Caruso
    • 2
  • Quirino Alves de Lima Neto
    • 3
  • Francisco Ferreira Duarte Junior
    • 3
  • Fabiana dos Santos Rando
    • 4
  • Edileusa Cristina Marques Gerhardt
    • 5
  • Maria Aparecida Fernandez
    • 3
  • Flávio Augusto Vicente Seixas
    • 1
    Email author
  1. 1.Department of TechnologyUniversidade Estadual de Maringá-UEMUmuaramaBrazil
  2. 2.Department of Physics, Instituto de BiociênciasLetras e Ciências Exatas-Universidade Estadual Paulista “Júlio de Mesquita Filho”São José do Rio PretoBrazil
  3. 3.Department of Biotechnology, Genetics and Cell BiologyUniversidade Estadual de MaringáMaringáBrazil
  4. 4.Center for Molecular, Structural and Functional Biology, CBM-Research Support Center ComplexUniversidade Estadual de MaringáMaringáBrazil
  5. 5.Department of BiochemistryUniversidade Federal do ParanáCuritibaBrazil

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