The effects of Ca2+ binding on the conformation of calbindin D28K

A nuclear magnetic resonance and microelectrospray mass spectrometry study

Ronald A. Venters, Linda M. Benson, Theodore A. Craig, Keriann H. Paul, David R. Kordys, Richele Thompson, Stephen Naylor, Rajiv Kumar, John Cavanagh

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Calbindin D28K is a six-EF-hand calcium-binding protein found in the brain, peripheral nervous system, kidney, and intestine. There is a paucity of information on the effects of calcium binding on calbindin D28K structure. To further examine the mechanism and structural consequences of calcium binding to calbindin D28K we performed detailed complementary heteronuclear NMR and microelectrospray mass spectrometry investigations of the calcium-induced conformational changes of calbindin D28K. The combined use of these two powerful analytical techniques clearly and very rapidly demonstrates the following: (i) apo-calbindin D28K has an ordered structure which changes to a notably different ordered conformation upon Ca2+ loading, (ii) calcium binding is a sequential process and not a simultaneous event, and (iii) EF-hands 1, 3, 4, and 5 take up Ca2+, whereas EF-hands 2 and 6 do not. Our results support the opinion that calbindin D28K has characteristics of both a calcium sensor and a buffer.

Original languageEnglish (US)
Pages (from-to)59-66
Number of pages8
JournalAnalytical Biochemistry
Volume317
Issue number1
DOIs
StatePublished - Jun 1 2003

Fingerprint

Calbindin 1
Mass spectrometry
Conformations
Mass Spectrometry
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
EF Hand Motifs
Calcium
Peptide Elongation Factor 2
Peptide Elongation Factor 1
Biomolecular Nuclear Magnetic Resonance
Calcium-Binding Proteins
Peripheral Nervous System
Neurology
Intestines
Brain
Buffers
Kidney
Sensors

Keywords

  • Calbindin D
  • Conformational response to calcium binding
  • EF-Hand

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

The effects of Ca2+ binding on the conformation of calbindin D28K : A nuclear magnetic resonance and microelectrospray mass spectrometry study. / Venters, Ronald A.; Benson, Linda M.; Craig, Theodore A.; Paul, Keriann H.; Kordys, David R.; Thompson, Richele; Naylor, Stephen; Kumar, Rajiv; Cavanagh, John.

In: Analytical Biochemistry, Vol. 317, No. 1, 01.06.2003, p. 59-66.

Research output: Contribution to journalArticle

Venters, Ronald A. ; Benson, Linda M. ; Craig, Theodore A. ; Paul, Keriann H. ; Kordys, David R. ; Thompson, Richele ; Naylor, Stephen ; Kumar, Rajiv ; Cavanagh, John. / The effects of Ca2+ binding on the conformation of calbindin D28K : A nuclear magnetic resonance and microelectrospray mass spectrometry study. In: Analytical Biochemistry. 2003 ; Vol. 317, No. 1. pp. 59-66.
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