Inter-α-inhibitor in calcium stones

Dawson, Caroline J.; Grover, Phulwinder K.; Kanellos, Jerry; Pham, Hung; Kupczyk, Gosia; Oates, A.; Ryall, Rosemary L.

doi:10.1042/cs0950187

“…Williams et al [24] compared 4 different methods for COM stone dissolution and found extractible proteins to range only from 0.18-0.55% of stone weight. Dawson et al [38] extracted calcium oxalate (8 stones) or calcium phosphate stones (2 stones) with 0.25M EDTA for 48h, and found variable amounts of protein; in some samples, protein bands were barely visible. Sugimoto et al [39] extracted stone proteins in saline by dialyzing against 0.1 M EDTA, and found protein levels were significantly increased in calcium phosphate (3 stones) over calcium oxalate stones, with an average extractible protein content of all stones of 1.6%.…”

Section: Discussionmentioning

confidence: 99%

Protein content of human apatite and brushite kidney stones: significant correlation with morphologic measures

Pramanik

¹

,

Asplin

²

,

Jackson

³

et al. 2008

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Apatite and brushite kidney stones share calcium and phosphate as their main inorganic components. We tested the hypothesis that these stone types differ in the organic matrix present in the stones. Intact stones were intensively analyzed by micro-computed tomography (micro CT) for both morphology (including the volume of voids, i.e., regions without mineral) and mineral type. In order to extract all proteins present in kidney stones in soluble form we developed a three-step extraction procedure using the ground stone powder. Apatite stones had significantly higher levels of total protein content and void volume compared to brushite stones. The void volume was highly correlated with the total protein contents in all stones (r2=0.61, P<0.0001), and brushite stones contained significantly fewer void regions and proteins than did apatite stones (3.2±4.5% voids for brushite vs. 10.8±11.2% for apatite, P<0.005; 4.1±1.6% protein for brushite vs. 6.0±2.4% for apatite, P<0.03). Morphological observations other than void volume did not correlate with protein content of stones, and neither did the presence or absence of minor mineral components. Our results show that protein content of brushite and apatite stones is higher than was previously thought, and also suggest that micro CT-visible void regions are related to the presence of protein.

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“…IαI was purified from Prothrombinex2-HT, a concentrate of human blood coagulation factors manufactured from human plasma by CSL Ltd. (Victoria, Australia), by gel permeation on a Superose 6 (5 cmi90 cm) column (Pharmacia, Uppsala, Sweden) with a citric acid\ phosphate buffer (pH 7.4) as described elsewhere [27]. All protein fractions were monitored at 280 nm and were analysed by SDS\PAGE and Western blotting (see below), and those fractions containing IαI were subjected to alkaline hydrolysis as detailed in [27], which dissociated IαI into various fragments.…”

Section: Purification Of Iαi Bikunin and H1 From Prothrombinex2-ht Bmentioning

confidence: 99%

“…However, the detection of H1 and H2 in normal human urine, calcium stones and CaOx crystals [27,28], and the presence of intact IαI in several normal urines [29], suggest that bikunin might not be the only IαI derivative with a possible involvement in stone formation. However, the detection of H1 and H2 in normal human urine, calcium stones and CaOx crystals [27,28], and the presence of intact IαI in several normal urines [29], suggest that bikunin might not be the only IαI derivative with a possible involvement in stone formation.…”

Section: Introductionmentioning

confidence: 99%

Effects of inter-α-inhibitor and several of its derivatives on calcium oxalate crystallization in vitro

Dean

¹

,

Kanellos

²

,

Pham

³

et al. 2000

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The bikunin peptide chain of the protease inhibitor inter-alpha-inhibitor (IalphaI) has been reported to be an inhibitor of calcium oxalate (CaOx) crystallization, and hence has been proposed as having a role in CaOx kidney stone formation. However, further experimental evidence is required to assess if fragments of IalphaI other than bikunin may play a role in the regulation of crystallization events in stone formation. The aim of the present study was to assess the effects of IalphaI and several of its derivatives on CaOx crystallization in a seeded inorganic system and to compare these effects with those of a known inhibitor of crystallization, prothrombin. IalphaI was purified from a preparation of human plasma and fragmented by alkaline hydrolysis, and two of its peptide chains, bikunin and heavy chain 1 (H1), were purified further by HPLC. Their purity was confirmed by SDS/PAGE. Using Coulter counter and [(14)C]oxalate analysis and scanning electron microscopy, IalphaI, its H1 chain and bikunin from urine and from plasma were shown to be relatively weak inhibitors of CaOx crystallization in vitro at expected physiological concentrations. It was concluded that members of the IalphaI family may not be as important in kidney stone formation as has been generally proposed, although further studies are required before a possible role for IalphaI and its fragments in stone formation can be unambiguously discounted.

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“…All protein fractions were monitored at 280 nm and were analysed by SDS\PAGE and Western blotting (see below), and those fractions containing IαI were subjected to alkaline hydrolysis as detailed in [27], which dissociated IαI into various fragments.…”

Section: Purification Of Iαi Bikunin and H1 From Prothrombinex2-ht Bmentioning

confidence: 99%

“…However, the detection of H1 and H2 in normal human urine, calcium stones and CaOx crystals [27,28], and the presence of intact IαI in several normal urines [29], suggest that bikunin might not be the only IαI derivative with a possible involvement in stone formation.…”

Section: Introductionmentioning

confidence: 99%

Effects of inter-α-inhibitor and several of its derivatives on calcium oxalate crystallization in vitro

DEAN¹,

KANELLOS²,

PHAM³

et al. 2000

Self Cite

View full text Add to dashboard Cite

The bikunin peptide chain of the protease inhibitor inter-alpha-inhibitor (IalphaI) has been reported to be an inhibitor of calcium oxalate (CaOx) crystallization, and hence has been proposed as having a role in CaOx kidney stone formation. However, further experimental evidence is required to assess if fragments of IalphaI other than bikunin may play a role in the regulation of crystallization events in stone formation. The aim of the present study was to assess the effects of IalphaI and several of its derivatives on CaOx crystallization in a seeded inorganic system and to compare these effects with those of a known inhibitor of crystallization, prothrombin. IalphaI was purified from a preparation of human plasma and fragmented by alkaline hydrolysis, and two of its peptide chains, bikunin and heavy chain 1 (H1), were purified further by HPLC. Their purity was confirmed by SDS/PAGE. Using Coulter counter and [(14)C]oxalate analysis and scanning electron microscopy, IalphaI, its H1 chain and bikunin from urine and from plasma were shown to be relatively weak inhibitors of CaOx crystallization in vitro at expected physiological concentrations. It was concluded that members of the IalphaI family may not be as important in kidney stone formation as has been generally proposed, although further studies are required before a possible role for IalphaI and its fragments in stone formation can be unambiguously discounted.

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Inter-α-inhibitor in calcium stones

Cited by 16 publications

References 35 publications

Protein content of human apatite and brushite kidney stones: significant correlation with morphologic measures

Protein content of human apatite and brushite kidney stones: significant correlation with morphologic measures

Effects of inter-α-inhibitor and several of its derivatives on calcium oxalate crystallization in vitro

Effects of inter-α-inhibitor and several of its derivatives on calcium oxalate crystallization in vitro

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