Ca2+-Binding domain VI of rat calpain is a homodimer in solution: Hydrodynamic, crystallization and preliminary X-ray diffraction studies

Helen Blanchard; Yunge Li; Miroslaw Cygler; Cyril M. Kay; J. Simon C. Arthur; Peter L. Davies; John S. Elce

doi:10.1002/pro.5560050317

Ca²⁺-Binding domain VI of rat calpain is a homodimer in solution: Hydrodynamic, crystallization and preliminary X-ray diffraction studies

Helen Blanchard
, Yunge Li
, Miroslaw Cygler (Lead / Corresponding author)
, Cyril M. Kay
, J. Simon C. Arthur
, Peter L. Davies
, John S. Elce

Cell Signalling and Immunology

Research output: Contribution to journal › Article › peer-review

32 Citations (Scopus)

Abstract

The 21-kDa calcium-binding domain (VI) of the small subunit of rat calpain II has been expressed in Escherichia coli, purified, and crystallized. Two orthorhombic crystal forms have been obtained: space group P2₁2₁2₁ with a = 50.3, b = 56.5, c = 141.3 Å; and space group C222₁ with a = 69.4, b = 73.9, c = 157.4 Å. Diffraction data have been collected to 2.4 Å. Sedimentation equilibrium, dynamic light scattering, and gel-permeation chromatography indicate that domain VI exists as a homodimer in solution. In accordance with the protein's behavior in solution, each crystal form contains two molecules per asymmetric unit. Screening for heavy-atom derivatives is in progress. To decrease the sensitivity to mercurials and to aid in the search for useful derivatives, Cys-to-Ser mutants have been prepared, expressed, and crystallized.

Original language	English
Pages (from-to)	535-537
Number of pages	3
Journal	Protein Science
Volume	5
Issue number	3
DOIs	https://doi.org/10.1002/pro.5560050317
Publication status	Published - Mar 1996

Keywords

Calcium-binding domain
Calpain
Crystallization
Homodimer
Smalt subunit

ASJC Scopus subject areas

Biochemistry
Molecular Biology

Access to Document

10.1002/pro.5560050317

Cite this

@article{38d78c9fbc404f35b2f2268b0cfd569b,

title = "Ca2+-Binding domain VI of rat calpain is a homodimer in solution: Hydrodynamic, crystallization and preliminary X-ray diffraction studies",

abstract = "The 21-kDa calcium-binding domain (VI) of the small subunit of rat calpain II has been expressed in Escherichia coli, purified, and crystallized. Two orthorhombic crystal forms have been obtained: space group P212121 with a = 50.3, b = 56.5, c = 141.3 {\AA}; and space group C2221 with a = 69.4, b = 73.9, c = 157.4 {\AA}. Diffraction data have been collected to 2.4 {\AA}. Sedimentation equilibrium, dynamic light scattering, and gel-permeation chromatography indicate that domain VI exists as a homodimer in solution. In accordance with the protein's behavior in solution, each crystal form contains two molecules per asymmetric unit. Screening for heavy-atom derivatives is in progress. To decrease the sensitivity to mercurials and to aid in the search for useful derivatives, Cys-to-Ser mutants have been prepared, expressed, and crystallized.",

keywords = "Calcium-binding domain, Calpain, Crystallization, Homodimer, Smalt subunit",

author = "Helen Blanchard and Yunge Li and Miroslaw Cygler and Kay, \{Cyril M.\} and Arthur, \{J. Simon C.\} and Davies, \{Peter L.\} and Elce, \{John S.\}",

year = "1996",

month = mar,

doi = "10.1002/pro.5560050317",

language = "English",

volume = "5",

pages = "535--537",

journal = "Protein Science",

issn = "0961-8368",

publisher = "Wiley",

number = "3",

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TY - JOUR

T1 - Ca2+-Binding domain VI of rat calpain is a homodimer in solution

T2 - Hydrodynamic, crystallization and preliminary X-ray diffraction studies

AU - Blanchard, Helen

AU - Li, Yunge

AU - Cygler, Miroslaw

AU - Kay, Cyril M.

AU - Arthur, J. Simon C.

AU - Davies, Peter L.

AU - Elce, John S.

PY - 1996/3

Y1 - 1996/3

N2 - The 21-kDa calcium-binding domain (VI) of the small subunit of rat calpain II has been expressed in Escherichia coli, purified, and crystallized. Two orthorhombic crystal forms have been obtained: space group P212121 with a = 50.3, b = 56.5, c = 141.3 Å; and space group C2221 with a = 69.4, b = 73.9, c = 157.4 Å. Diffraction data have been collected to 2.4 Å. Sedimentation equilibrium, dynamic light scattering, and gel-permeation chromatography indicate that domain VI exists as a homodimer in solution. In accordance with the protein's behavior in solution, each crystal form contains two molecules per asymmetric unit. Screening for heavy-atom derivatives is in progress. To decrease the sensitivity to mercurials and to aid in the search for useful derivatives, Cys-to-Ser mutants have been prepared, expressed, and crystallized.

AB - The 21-kDa calcium-binding domain (VI) of the small subunit of rat calpain II has been expressed in Escherichia coli, purified, and crystallized. Two orthorhombic crystal forms have been obtained: space group P212121 with a = 50.3, b = 56.5, c = 141.3 Å; and space group C2221 with a = 69.4, b = 73.9, c = 157.4 Å. Diffraction data have been collected to 2.4 Å. Sedimentation equilibrium, dynamic light scattering, and gel-permeation chromatography indicate that domain VI exists as a homodimer in solution. In accordance with the protein's behavior in solution, each crystal form contains two molecules per asymmetric unit. Screening for heavy-atom derivatives is in progress. To decrease the sensitivity to mercurials and to aid in the search for useful derivatives, Cys-to-Ser mutants have been prepared, expressed, and crystallized.

KW - Calcium-binding domain

KW - Calpain

KW - Crystallization

KW - Homodimer

KW - Smalt subunit

UR - https://www.scopus.com/pages/publications/0029670455

U2 - 10.1002/pro.5560050317

DO - 10.1002/pro.5560050317

M3 - Article

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AN - SCOPUS:0029670455

SN - 0961-8368

VL - 5

SP - 535

EP - 537

JO - Protein Science

JF - Protein Science

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