1H NMR assignments of apo calcyclin and comparative structural analysis with calbindin d(9k) and s 100β
Potts, Barbara C.M. ; Carlström, Göran LU
; Okazaki, Katsuo
; Hidaka, Hiroyoshi
and Chazin, Walter J.
(1996)
In Protein Science
5(11).
p.2162-2174
- Abstract
The homodimeric S100 protein calcyclin has been studied in the apo state by two-dimensional 1H NMR spectroscopy. Using a combination of scalar correlation and NOE experiments, sequence-specific 1H NMR assignments were obtained for all but one backbone and >90% of the side-chain resonances. To our knowledge, the 2 x 90 residue (20 kDa) calcyclin dimer is the largest protein system for which such complete assignments have been made by purely homonuclear methods. Sequential and medium-range NOEs and slowly exchanging backbone amide protons identified directly the four helices and the short antiparallel β-type interaction between the two binding loops that comprise each subunit of the dimer. Further analysis of NOEs... (More)
The homodimeric S100 protein calcyclin has been studied in the apo state by two-dimensional 1H NMR spectroscopy. Using a combination of scalar correlation and NOE experiments, sequence-specific 1H NMR assignments were obtained for all but one backbone and >90% of the side-chain resonances. To our knowledge, the 2 x 90 residue (20 kDa) calcyclin dimer is the largest protein system for which such complete assignments have been made by purely homonuclear methods. Sequential and medium-range NOEs and slowly exchanging backbone amide protons identified directly the four helices and the short antiparallel β-type interaction between the two binding loops that comprise each subunit of the dimer. Further analysis of NOEs enabled the unambiguous assignment of 556 intrasubunit distance constraints, 24 intrasubunit hydrogen bonding constraints, and 2 x 26 intersubunit distance constraints. The conformation of the monomer subunit was refined by distance geometry and restrained molecular dynamics calculations using the intrasubunit constraints only. Calculation of the dimer structure starting from this conformational ensemble has been reported elsewhere. The extent of structural homology among the apo calcyclin subunit, the monomer subunit of apo S100β, and monomeric apo calbindin D(9k) has been examined in detail by comparing 1H NMR chemical shifts and secondary structures. This analysis was extended to a comprehensive comparison of the three-dimensional structures of the calcyclin monomer subunit and calbindin D(9k), which revealed greater similarity in the packing of their hydrophobic cores than was anticipated previously. Together, these results support the hypothesis that all members of the S100 family have similar core structures and similar modes of dimerization. Analysis of the amphiphilicity of Helix IV is used to explain why calbindin D(9k) is monomeric, but full-length S100 proteins form homodimers.
(Less)
- author
- Potts, Barbara C.M.
; Carlström, Göran
LU
; Okazaki, Katsuo
; Hidaka, Hiroyoshi
and Chazin, Walter J.
- organization
- publishing date
- 1996-01-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- calcyclin, chemical shift, EF-hand calcium-binding protein, homology, hydrophobic core, S100 protein
- in
- Protein Science
- volume
- 5
- issue
- 11
- pages
- 13 pages
- publisher
- The Protein Society
- external identifiers
-
- pmid:8931135
- scopus:0029658223
- ISSN
- 0961-8368
- DOI
- 10.1002/pro.5560051103
- language
- English
- LU publication?
- yes
- id
- ef0b11f9-0174-48bd-993b-c08c587b7631
- date added to LUP
- 2019-07-25 21:37:21
- date last changed
- 2024-01-01 16:52:44
@article{ef0b11f9-0174-48bd-993b-c08c587b7631,
abstract = {{<p>The homodimeric S100 protein calcyclin has been studied in the apo state by two-dimensional <sup>1</sup>H NMR spectroscopy. Using a combination of scalar correlation and NOE experiments, sequence-specific <sup>1</sup>H NMR assignments were obtained for all but one backbone and >90% of the side-chain resonances. To our knowledge, the 2 x 90 residue (20 kDa) calcyclin dimer is the largest protein system for which such complete assignments have been made by purely homonuclear methods. Sequential and medium-range NOEs and slowly exchanging backbone amide protons identified directly the four helices and the short antiparallel β-type interaction between the two binding loops that comprise each subunit of the dimer. Further analysis of NOEs enabled the unambiguous assignment of 556 intrasubunit distance constraints, 24 intrasubunit hydrogen bonding constraints, and 2 x 26 intersubunit distance constraints. The conformation of the monomer subunit was refined by distance geometry and restrained molecular dynamics calculations using the intrasubunit constraints only. Calculation of the dimer structure starting from this conformational ensemble has been reported elsewhere. The extent of structural homology among the apo calcyclin subunit, the monomer subunit of apo S100β, and monomeric apo calbindin D(9k) has been examined in detail by comparing <sup>1</sup>H NMR chemical shifts and secondary structures. This analysis was extended to a comprehensive comparison of the three-dimensional structures of the calcyclin monomer subunit and calbindin D(9k), which revealed greater similarity in the packing of their hydrophobic cores than was anticipated previously. Together, these results support the hypothesis that all members of the S100 family have similar core structures and similar modes of dimerization. Analysis of the amphiphilicity of Helix IV is used to explain why calbindin D(9k) is monomeric, but full-length S100 proteins form homodimers.</p>}},
author = {{Potts, Barbara C.M. and Carlström, Göran and Okazaki, Katsuo and Hidaka, Hiroyoshi and Chazin, Walter J.}},
issn = {{0961-8368}},
keywords = {{calcyclin; chemical shift; EF-hand calcium-binding protein; homology; hydrophobic core; S100 protein}},
language = {{eng}},
month = {{01}},
number = {{11}},
pages = {{2162--2174}},
publisher = {{The Protein Society}},
series = {{Protein Science}},
title = {{<sup>1</sup>H NMR assignments of apo calcyclin and comparative structural analysis with calbindin d(9k) and s 100β}},
url = {{http://dx.doi.org/10.1002/pro.5560051103}},
doi = {{10.1002/pro.5560051103}},
volume = {{5}},
year = {{1996}},
}