Structure and properties of heavy-metal binding protein from
Agaricus bisporus. A case study on utilizing X-ray
crystallography and different biophysical techniques for
elucidating protein function

a 2017

Structure and properties of heavy-metal binding protein from Agaricus bisporus. A case study on utilizing X-ray crystallography and different biophysical techniques for elucidating protein function

KOMÁREK, Jan, Eva KAVKOVÁ, Josef HOUSER, Aneta HORÁČKOVÁ, Jitka ŽDÁNSKÁ et. al.

Basic information

Original name

Structure and properties of heavy-metal binding protein from Agaricus bisporus. A case study on utilizing X-ray crystallography and different biophysical techniques for elucidating protein function

Authors

KOMÁREK, Jan, Eva KAVKOVÁ, Josef HOUSER, Aneta HORÁČKOVÁ, Jitka ŽDÁNSKÁ and Michaela WIMMEROVÁ

Edition

23rd International Analytical Ultracentrifugation Workshop and Symposium, 2017

Other information

Language

English

Type of outcome

Konferenční abstrakt

Field of Study

10600 1.6 Biological sciences

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

Organization unit

Central European Institute of Technology

Keywords in English

metal-binding proteins; Agaricus bisporus

Změněno: 24/1/2018 09:32, Mgr. Jan Komárek, Ph.D.

Abstract

V originále

The white button mushroom Agaricus bisporus is one of the most widely cultivated mushrooms in the world and an important component of human diet. Apart from its high nutritional value, it has a significant ecological role (e.g. involvement in the decay of a leaf litter). Here we report the structure-functional characterization of 45 kDa dimeric protein from A. bisporus. The protein shares no detectable sequence homology to any protein of known function and it is the first characterized member of the protein family. The cDNA sequence was amplified by the reverse transcription PCR and the protein was expressed in E. coli in a recombinant form. The X-ray structure was solved to a resolution of 1.6 A and it showed an elongated helical bundle structure with rather low structural similarity to other proteins. Using different biophysical techniques (AUC, CD, DSF) it was shown that the protein displays high thermostability and is stable over a wide range of pH. Finally, with DSC and ITC we identified and characterized its ability to bind divalent heavy metal ions (nickel, zinc, cadmium and cobalt), which might suggest its possible role in homeostasis, regulational pathways or sequestration and removal of heavy metals.

Links

CZ.02.1.01/0.0/0.0/16_013/0001776, interní kód MU
(CEP code: EF16_013/0001776)

Name: CIISB - Česká infrastruktura pro integrativní strukturní biologii pro lidské zdraví (Acronym: CIISB4HEALTH)

Investor: Ministry of Education, Youth and Sports of the CR, Czech Infrastructure for Integrative Structural Biology for Human Health, Priority axis 1: Strengthening capacities for high-quality research

Citovat

KOMÁREK, Jan, Eva KAVKOVÁ, Josef HOUSER, Aneta HORÁČKOVÁ, Jitka ŽDÁNSKÁ and Michaela WIMMEROVÁ. Structure and properties of heavy-metal binding protein from Agaricus bisporus. A case study on utilizing X-ray crystallography and different biophysical techniques for elucidating protein function. In 23rd International Analytical Ultracentrifugation Workshop and Symposium. 2017.

@proceedings{1404172,
   author = {Komárek, Jan and Kavková, Eva and Houser, Josef and Horáčková, Aneta and Ždánská, Jitka and Wimmerová, Michaela},
   booktitle = {23rd International Analytical Ultracentrifugation Workshop and Symposium},
   keywords = {metal-binding proteins; Agaricus bisporus},
   language = {eng},
   title = {Structure and properties of heavy-metal binding protein from Agaricus bisporus. A case study on utilizing X-ray crystallography and different biophysical techniques for elucidating protein function},
   year = {2017}
}

TY  - CONF
ID  - 1404172
AU  - Komárek, Jan - Kavková, Eva - Houser, Josef - Horáčková, Aneta - Ždánská, Jitka - Wimmerová, Michaela
PY  - 2017
TI  - Structure and properties of heavy-metal binding protein from Agaricus bisporus. A case study on utilizing X-ray crystallography and different biophysical techniques for elucidating protein function
KW  - metal-binding proteins
KW  - Agaricus bisporus
N2  - The white button mushroom Agaricus bisporus is one of the most widely cultivated mushrooms in the world and an important component of human diet. Apart from its high nutritional value, it has a significant ecological role (e.g. involvement in the decay of a leaf litter). Here we report the structure-functional characterization of 45 kDa dimeric protein from A. bisporus. The protein shares no detectable sequence homology to any protein of known function and it is the first characterized member of the protein family. The cDNA sequence was amplified by the reverse transcription PCR and the protein was expressed in E. coli in a recombinant form. The X-ray structure was solved to a resolution of 1.6 A and it showed an elongated helical bundle structure with rather low structural similarity to other proteins. Using different biophysical techniques (AUC, CD, DSF) it was shown that the protein displays high thermostability and is stable over a wide range of pH. Finally, with DSC and ITC we identified and characterized its ability to bind divalent heavy metal ions (nickel, zinc, cadmium and cobalt), which might suggest its possible role in homeostasis, regulational pathways or sequestration and removal of heavy metals.
ER  -

KOMÁREK, Jan, Eva KAVKOVÁ, Josef HOUSER, Aneta HORÁČKOVÁ, Jitka ŽDÁNSKÁ and Michaela WIMMEROVÁ. Structure and properties of heavy-metal binding protein from Agaricus bisporus. A case study on utilizing X-ray crystallography and different biophysical techniques for elucidating protein function. In \textit{23rd International Analytical Ultracentrifugation Workshop and Symposium}. 2017.

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