Insertion of Heme b into the Structure of the Cys34-Carbamidomethylated Human Lipocalin a1-Microglobulin: Formation of a [(Heme)2(a1-Microglobulin)]3 Complex
(2012) In ChemBioChem 13(6). p.879-887- Abstract
- a1-Microglobulin (a1m) is a 26 kDa plasma and tissue protein belonging to the lipocalin protein family. Previous investigations indicate that the protein interacts with heme and suggest that it has a function in heme metabolism. However, detailed characterizations of the a1mheme interactions are lacking. Here, we report for the first time the preparation and analysis of a stable a1mheme complex upon carbamidomethylation of the reactive Cys34 by using recombinantly expressed human a1m. Analytical size-exclusion chromatography coupled with a diode-array absorbance spectrophotometry demonstrates that at first an a1mheme monomer is formed. Subsequently, a second heme triggers oligomerization that leads to trimerization. The resulting... (More)
- a1-Microglobulin (a1m) is a 26 kDa plasma and tissue protein belonging to the lipocalin protein family. Previous investigations indicate that the protein interacts with heme and suggest that it has a function in heme metabolism. However, detailed characterizations of the a1mheme interactions are lacking. Here, we report for the first time the preparation and analysis of a stable a1mheme complex upon carbamidomethylation of the reactive Cys34 by using recombinantly expressed human a1m. Analytical size-exclusion chromatography coupled with a diode-array absorbance spectrophotometry demonstrates that at first an a1mheme monomer is formed. Subsequently, a second heme triggers oligomerization that leads to trimerization. The resulting (a1m[heme]2)3 complex was characterized by resonance Raman and EPR spectroscopy, which support the presence of two ferrihemes, thus indicating an unusual spin-state admixed ground state with S=3/2, 5/2. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2571583
- author
- Siebel, Judith F. ; Kosinsky, Robyn L. ; Åkerström, Bo LU and Knipp, Markus
- organization
- publishing date
- 2012
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- heme proteins, homotrimers, lipocalin, metalloproteins, microglobulins, spin admixture
- in
- ChemBioChem
- volume
- 13
- issue
- 6
- pages
- 879 - 887
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- wos:000302609100018
- scopus:84859597177
- ISSN
- 1439-4227
- DOI
- 10.1002/cbic.201100808
- language
- English
- LU publication?
- yes
- id
- 9010a33b-8c6d-485c-9698-4cebf013e68a (old id 2571583)
- date added to LUP
- 2016-04-01 09:56:43
- date last changed
- 2022-01-25 18:15:21
@article{9010a33b-8c6d-485c-9698-4cebf013e68a, abstract = {{a1-Microglobulin (a1m) is a 26 kDa plasma and tissue protein belonging to the lipocalin protein family. Previous investigations indicate that the protein interacts with heme and suggest that it has a function in heme metabolism. However, detailed characterizations of the a1mheme interactions are lacking. Here, we report for the first time the preparation and analysis of a stable a1mheme complex upon carbamidomethylation of the reactive Cys34 by using recombinantly expressed human a1m. Analytical size-exclusion chromatography coupled with a diode-array absorbance spectrophotometry demonstrates that at first an a1mheme monomer is formed. Subsequently, a second heme triggers oligomerization that leads to trimerization. The resulting (a1m[heme]2)3 complex was characterized by resonance Raman and EPR spectroscopy, which support the presence of two ferrihemes, thus indicating an unusual spin-state admixed ground state with S=3/2, 5/2.}}, author = {{Siebel, Judith F. and Kosinsky, Robyn L. and Åkerström, Bo and Knipp, Markus}}, issn = {{1439-4227}}, keywords = {{heme proteins; homotrimers; lipocalin; metalloproteins; microglobulins; spin admixture}}, language = {{eng}}, number = {{6}}, pages = {{879--887}}, publisher = {{John Wiley & Sons Inc.}}, series = {{ChemBioChem}}, title = {{Insertion of Heme b into the Structure of the Cys34-Carbamidomethylated Human Lipocalin a1-Microglobulin: Formation of a [(Heme)2(a1-Microglobulin)]3 Complex}}, url = {{http://dx.doi.org/10.1002/cbic.201100808}}, doi = {{10.1002/cbic.201100808}}, volume = {{13}}, year = {{2012}}, }