The isolation and identification of α1-antichymotrypsin as a DNA-binding protein from human serum

Siddiqui, Anwar Ali; Hughes, Alun; Davies, R. Jeremy H.; Hill, John

doi:10.1016/s0006-291x(80)80099-4

Cited by 15 publications

(6 citation statements)

References 15 publications

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“…(26) of this data suggests that HDP/LDH binds ss DNA with an affinity that is between 106 and 107 M-1 in 0.05 M NaCl (27), which compares with a value of 1014 M-1 for E. coli SSB under the same conditions (unpublished data). In addition, several other proteins, including the P-1 protein (protocollagen precursor) from fibroblasts (28), a-1-antichymotrypsin (29), and three serum proteins involved in complement activation: C3DP (30), factor B (31), and f31H, (32) that apparently have no in vivo role in DNA metabolism nonetheless bind to ss DNA-cellulose. Taken together, these data indicate that some care needs to be exercised in ascribing in vivo functions simply on the basis of binding to ss DNA-cellulose (2) or acting as a helix-destabilizing protein in vitro.…”

Section: Discussionmentioning

confidence: 99%

Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5.

Williams¹,

Reddigari²,

Patel³

1985

Proc. Natl. Acad. Sci. U.S.A.

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A rat liver DNA helix-destabilizing protein (HDP) that has previously been proposed to play a role in transcription has been identified as M chain lactate dehydrogenase (LDH-5; L-lactate:NAD+ oxidoreductase, EC 1.1.1.27). Tryptic peptides accounting for 157 amino acids in the rat liver HDP have been characterized and then matched to the published sequence for the M chain of porcine LDH. Based on amino acid compositions and direct solid-phase protein sequencing, at least 148 of the 157 residues that were compared are identical in both proteins. In addition, both porcine LDH and the rat liver HDP have blocked amino termini and similar amino acid compositions and molecular weights. Rat liver HDP and LDH-5 that were purified to molecular homogeneity had similar specific activities in both single-stranded DNA (ss DNA) binding and LDH assays. HPLC tryptic peptide maps were also identical for both the rat liver HDP and LDH proteins. Since preincubation of HDP in NADH prevents its binding to ss DNA, both NADH and ss DNA may be binding at the same site. Further support for this latter idea derives from chemicalmodification studies which demonstrate that tyrosine-238, which is located near the coenzyme binding site of LDH, seems to be essential for the ability of HDP to bind ss DNA. These results indicate caution in ascribing in vivo roles solely on the basis of binding to ss DNA. Alternatively, they suggest that a single protein may play more than one biological role.

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Section: Discussionmentioning

confidence: 99%

Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5.

Williams¹,

Reddigari²,

Patel³

1985

Proc. Natl. Acad. Sci. U.S.A.

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“…No homozygotes have yet been identified. a,-Antichymotrypsin has been shown to be identical to the protein termed 64DP [9] and has a high affinity for DNAcellulose.It has been claimed that a,-antichymotrypsin is a component of the brain amyloid deposits in Alzheimer's disease [lo] but we recently showed 1111 that no distinction can be made between extracts of 14 control brains and extracts of 12 Alzheimer brains with regard to the amount of al-antichymotrypsin.The complete amino acid sequence of al-antichymotrypsin deduced from the nucleotide sequence [I21 reveals four poten- a,-Antichymotrypsin purified from the serum of a single healthy donor, in order to avoid possible genetic heterogeneity, was separated by affinity chromatography into three fractions on a column of concanavalin A (ConA)-linked to Sepharose [14]: a pass-through fraction, a retarded fraction and a bound fraction which was eluted from the column by the addition of methyl a-D-glucoside to the buffer. Our preliminary results [I41 suggest that these three fractions carry glycans with decreasing degrees of branching from the ConA pass-through form to the ConA-bound form.…”

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confidence: 99%

“…No homozygotes have yet been identified. a,-Antichymotrypsin has been shown to be identical to the protein termed 64DP [9] and has a high affinity for DNAcellulose.…”

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confidence: 99%

Structure determination of the glycans of human‐serum α₁‐antichymotrypsin using ¹H‐NMR spectroscopy and deglycosylation by N‐glycanase

Laine

Hachulla

Strecker

et al. 1991

European Journal of Biochemistry

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a,-Antichymotrypsin purified from normal human serum was separated by affinity chromatography into three microheterogeneous forms on a concanavalin-A -Sepharose column: a pass-through (peak I), a retarded (peak 2) and a bound form (peaks 3 + 4). For each form the asparagine-linked carbohydrate chains were liberated as oligosaccharides by hydrazinolysis, submitted to reduction with NaBH4 after re-N-acetylation and further separated by affinity chromatography on a concanavalin-A -Sepharose column. The complete primary structure of the glycans was determined by high-resolution 'H-NMR spectroscopy. The results indicated the presence of disialyl diantennary and of trisialyl triantennary type glycanic structures, the latter being accompanied by traces of disialylated triantennary oligosaccharide. The N-glycanase was used for the deglycosylation of the unfractionated ccl-antichymotrypsin; the successive removal of the N-linked complex-type oligosaccharide side chains of u,-antichymotrypsin was studied in the presence of detergents. From these experiments it is concluded that alantichymotrypsin carries four oligosaccharide side chains. Moreover our results show that the peak l contains four triantennary glycans, the peak 2 three triantennary and one diantennary glycans while the bound peaks 3 + 4 possess, on average, about one triantennary and three diantennary glycans per molecule. Since we showed that the peak 4 contains mostly diantennary glycans, it can be deduced that in peak 3 there are molecules carrying two triantennary and two diantennary glycans and others carrying one triantennary and three diantennary glycans.Human a,-antichymotrypsin is a plasma glycoprotein with a relative molecular mass ( M J of approximately 58000 and a carbohydrate content of 24% [l]. It belongs to the superfamily of serpins (serine proteinase inhibitors) [2]. In addition to Creactive protein and serum amyloid A protein, cr,-antichymotrypsin is one of the major acute-phase proteins [3] since its serum concentration increases more than fourfold within a few hours in response to an inflammatory stimulus [4].The precise biological function of a,-antichymotrypsin has not been yet determined. It is known to inhibit chymotrypsinlike proteases [5 -71 by formation of equimolar complexes which, however, dissociate with time. Eriksson et al.[8] recently described four families with inherited partial al-antichymotrypsin deficiency in which heterozygotes have 50% of normal plasma levels. No homozygotes have yet been identified. a,-Antichymotrypsin has been shown to be identical to the protein termed 64DP [9] and has a high affinity for DNAcellulose.It has been claimed that a,-antichymotrypsin is a component of the brain amyloid deposits in Alzheimer's disease [lo] but we recently showed 1111 that no distinction can be made between extracts of 14 control brains and extracts of 12 Alzheimer brains with regard to the amount of al-antichymotrypsin.The complete amino acid sequence of al-antichymotrypsin deduced from the nucleotide sequence [I21 reveals ...

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“…A further complication in the identification of functionally homologous proteins in eukaryotes is that numerous proteins, such as dehydrogenases (6, 7), a protocollagen precursor (8), serum proteins involved in complement activation (9, 10), a1-antichymotrypsin (11), and several other plasma proteins (12), that apparently have no in vivo role in DNA metabolism nonetheless bind reasonably well to ss DNA-cellulose. This affinity support is usually the first chromatographic step used for purifying ss DNA binding proteins.…”

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Amino acid sequence of the UP1 calf thymus helix-destabilizing protein and its homology to an analogous protein from mouse myeloma.

Williams¹,

Stone²,

LoPresti³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

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The isolation and identification of α1-antichymotrypsin as a DNA-binding protein from human serum

Cited by 15 publications

References 15 publications

Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5.

Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5.

Structure determination of the glycans of human‐serum α₁‐antichymotrypsin using ¹H‐NMR spectroscopy and deglycosylation by N‐glycanase

Amino acid sequence of the UP1 calf thymus helix-destabilizing protein and its homology to an analogous protein from mouse myeloma.

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The isolation and identification of α1-antichymotrypsin as a DNA-binding protein from human serum

Cited by 15 publications

References 15 publications

Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5.

Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5.

Structure determination of the glycans of human‐serum α1‐antichymotrypsin using 1H‐NMR spectroscopy and deglycosylation by N‐glycanase

Amino acid sequence of the UP1 calf thymus helix-destabilizing protein and its homology to an analogous protein from mouse myeloma.

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Structure determination of the glycans of human‐serum α₁‐antichymotrypsin using ¹H‐NMR spectroscopy and deglycosylation by N‐glycanase