Isolation of plaice (Pleuronectes platessa) α1-microglobulin : Conservation of structure and chromophore
(1999) In BBA - Protein Structure and Molecular Enzymology 1430(2). p.222-233- Abstract
A cDNA coding for plaice (Pleuronectes platessa) α1-microglobulin (Leaver et al., 1994, Comp. Biochem. Physiol. 108B, 275-281) was expressed and purified from baculovirus-infected insect cells. Specific monoclonal antibodies were then prepared and used to isolate the protein from plaice liver and serum. Mature 28.5 kDa α1-microglobulin was found in both liver and serum. The protein consisted of an 184 amino acid peptide with a complex N-glycan in position Asn123, one intrachain disulfide bridge and a yellow-brown chromophore. Physicochemical characterization indicated a globular shape with a frictional ratio of 1.37, electrophoretic charge-heterogeneity and antiparallel β-sheet structure. A smaller,... (More)
A cDNA coding for plaice (Pleuronectes platessa) α1-microglobulin (Leaver et al., 1994, Comp. Biochem. Physiol. 108B, 275-281) was expressed and purified from baculovirus-infected insect cells. Specific monoclonal antibodies were then prepared and used to isolate the protein from plaice liver and serum. Mature 28.5 kDa α1-microglobulin was found in both liver and serum. The protein consisted of an 184 amino acid peptide with a complex N-glycan in position Asn123, one intrachain disulfide bridge and a yellow-brown chromophore. Physicochemical characterization indicated a globular shape with a frictional ratio of 1.37, electrophoretic charge-heterogeneity and antiparallel β-sheet structure. A smaller, incompletely glycosylated, yellow-brown α1-microglobulin as well as a 45 kDa precursor protein were also found in liver. The chromophore was found to be linked to α1-microglobulin intracellularly. Recombinant plaice α1-microglobulin isolated from insect cells had the same N-terminal sequence, globular shape and yellow-brown color as mature α1-microglobulin, but carried a smaller, fucosylated, non-sialylated N-glycan in the Asn123 position. The concentration of α1-microglobulin in plaice serum was 20 mg/l and it was found both as a 28.5 kDa component and as high molecular weight components. Thus, the size, shape, charge and color of plaice α1-microglobulin were similar to mammalian α1-microglobulin, indicating a high degree of structural conservation between fish and human α1-microglobulin. The monoclonal antibodies against plaice α1-microglobulin cross-reacted with human α1-microglobulin, emphasizing the structural similarity. Copyright (C) 1999 Elsevier Science B.V.
(Less)
- author
- Lindqvist, Annika LU and Åkerström, Bo LU
- organization
- publishing date
- 1999-03-19
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- α-Microglobulin, Baculovirus, cDNA, Glycosylation, Liver, Serum, Protein HC, Teleost
- in
- BBA - Protein Structure and Molecular Enzymology
- volume
- 1430
- issue
- 2
- pages
- 222 - 233
- publisher
- Elsevier
- external identifiers
-
- scopus:0344196928
- pmid:10082950
- ISSN
- 0167-4838
- DOI
- 10.1016/S0167-4838(99)00003-5
- language
- English
- LU publication?
- yes
- id
- 7f2e1f0c-6ff3-44b3-ad24-0f317240c6c6
- date added to LUP
- 2019-06-27 12:16:13
- date last changed
- 2024-01-01 13:37:26
@article{7f2e1f0c-6ff3-44b3-ad24-0f317240c6c6,
abstract = {{<p>A cDNA coding for plaice (Pleuronectes platessa) α<sub>1</sub>-microglobulin (Leaver et al., 1994, Comp. Biochem. Physiol. 108B, 275-281) was expressed and purified from baculovirus-infected insect cells. Specific monoclonal antibodies were then prepared and used to isolate the protein from plaice liver and serum. Mature 28.5 kDa α<sub>1</sub>-microglobulin was found in both liver and serum. The protein consisted of an 184 amino acid peptide with a complex N-glycan in position Asn<sup>123</sup>, one intrachain disulfide bridge and a yellow-brown chromophore. Physicochemical characterization indicated a globular shape with a frictional ratio of 1.37, electrophoretic charge-heterogeneity and antiparallel β-sheet structure. A smaller, incompletely glycosylated, yellow-brown α<sub>1</sub>-microglobulin as well as a 45 kDa precursor protein were also found in liver. The chromophore was found to be linked to α<sub>1</sub>-microglobulin intracellularly. Recombinant plaice α<sub>1</sub>-microglobulin isolated from insect cells had the same N-terminal sequence, globular shape and yellow-brown color as mature α<sub>1</sub>-microglobulin, but carried a smaller, fucosylated, non-sialylated N-glycan in the Asn<sup>123</sup> position. The concentration of α<sub>1</sub>-microglobulin in plaice serum was 20 mg/l and it was found both as a 28.5 kDa component and as high molecular weight components. Thus, the size, shape, charge and color of plaice α<sub>1</sub>-microglobulin were similar to mammalian α<sub>1</sub>-microglobulin, indicating a high degree of structural conservation between fish and human α<sub>1</sub>-microglobulin. The monoclonal antibodies against plaice α<sub>1</sub>-microglobulin cross-reacted with human α<sub>1</sub>-microglobulin, emphasizing the structural similarity. Copyright (C) 1999 Elsevier Science B.V.</p>}},
author = {{Lindqvist, Annika and Åkerström, Bo}},
issn = {{0167-4838}},
keywords = {{α-Microglobulin; Baculovirus; cDNA; Glycosylation; Liver, Serum; Protein HC; Teleost}},
language = {{eng}},
month = {{03}},
number = {{2}},
pages = {{222--233}},
publisher = {{Elsevier}},
series = {{BBA - Protein Structure and Molecular Enzymology}},
title = {{Isolation of plaice (Pleuronectes platessa) α<sub>1</sub>-microglobulin : Conservation of structure and chromophore}},
url = {{http://dx.doi.org/10.1016/S0167-4838(99)00003-5}},
doi = {{10.1016/S0167-4838(99)00003-5}},
volume = {{1430}},
year = {{1999}},
}