Studies on the structure of heparin

Durant, G. J.; Hendrickson, H. R.; Montgomery, Rex

doi:10.1016/0003-9861(62)90289-8

Cited by 34 publications

(8 citation statements)

References 24 publications

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“…The glycosaminoglycans and their derivatives used as acceptor substrates for sulfotransferases were prepared according to the following references: heparan sulfate [2], Ndesulfoheparan sulfate [24], N,O-desulfoheparan sulfate [25,26], N-desulfo-N-[I -14C]acetylheparan sulfate [27], chondroitin 4/6-sulfate [28] and chondroitin [29]. N,O-desulfoheparan sulfate oligosaccharides were obtained after degradation of N,O-desulfoheparan sulfate with nitrous acid according to [15] with the modification that the degradation period was reduced to 10 rnin and the reaction was stopped by addition of 1 g urea/100 mg N,O-desulfoheparan sulfate.…”

Section: Substratesmentioning

confidence: 99%

“…It was identified on the basis of its resistance to chondroitin ABC lyase, its susceptibility to heparitinase, its sensitivity to nitrous acid and the presence of glucosamine as the only hexosamine. Mild acid hydrolysis of this heparansulfate [24] produced an N-desulfoheparan sulfate (relative molecular mass 16000 ; with a residual 0-sulfate content of 0.27 mol/mol disaccharide unit) which was used as the standard substrate for Ndesulfoheparan sulfate sulfotransferase.…”

Section: Substratesmentioning

confidence: 99%

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Purification and characterization of 3′‐phosphoadenylylsulfate: N‐desulfoheparan sulfate sulfotransferase from arterial tissue

Göhler

Niemann

Buddecke

1984

European Journal of Biochemistry

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1. A 3'-phosphoadenylsulfate : N-desulfoheparan sulfate sulfotransferase (EC 2.8.2.12) was purified 450-fold from the microsomal fraction of calf arterial tissue and separated from 3'-phosphoadenylylsulfate :chondroitin sulfotransferase (EC 2.8.2.5) activity.The enzyme has optimal activity at neutral pH, requires divalent cations (Mn", MgZ+, Ca") for maximal activity and exhibits specificity towards N-desulfoheparan sulfate, N,O-desulfoheparan sulfate and oligosaccharides derived therefrom. N,O-desulfoheparan sulfate tetrasaccharides serve as acceptor substrates only if the nonreducing terminus is occupied by glucuronic acid (not iduronic acid).2. Heparan sulfate is a major component of arterial tissue [1,2] and arterial smooth muscle cells [3,4], but has also been identified in various other mammalian tissues [5,6]. Heparan sulfate structurally resembles heparin, but in general heparan sulfate preparations are less sulfated and have a lower iduronic acid/glucuronic acid ratio than heparin. In its native form heparan sulfate is found covalently linked to a protein core as proteoglycan [l] and is an ubiquitous cell surface component [7,8] (and references cited therein), while heparin is largely stored as glycosaminoglycan intracellularly in the granules of mast cells (for review see [6] Enzymes. N-Acetylglucosamine deacetylase, 2-acetamido-2-deOXy-D-ghCOSe amidohydrolase (EC 3.5.1.33) ; desulfoheparan sulfotransferase, 3'-phosphoadenylylsulfate:N-desulfoheparin N-sulfotransferase (EC 2.8.2.8); heparitin sulfotransferase, 3-phosphoadenylylsulfate heparitin N-sulfotransferase (EC 2.8.2.1 2); j-glucuronidase, b-D-glucuronide glucuronosohydrolase (EC 3.2.1.31); heparitin sulfate lyase (EC 4.2.2.8); phospholipase A,, phosphatide 2-acylhydrolase (EC 3.1.1.4).Nomenclature. Heparan sulfate (heparitin sulfate) is a sulfated polymer consisting of alternating residues of uronic acid (glucuronic acid or iduronic acid) and glucosamine. It contains N-sulfated as well N-acetylated glucosamine residues and 0-sulfate groups [26]. Ndesulfoheparan sulfate is the chemically N-desulfated derivative of The biosynthesis of heparin has been studied extensively using cell-free systems based on mouse mastocytoma microsoma1 fractions [13 -211. The initially formed carbohydrate polymer undergoes a series of modifications involving Ndeacetylation, N-sulfation, epimerisation of glucuronic acid to iduronic acid and finally 0-sulfation in an ordered progress of events (for review see [22]), but further purification of the participating sulfotransferases has not been attempted.The present paper describes the purification and partial characterization of a 3'-phosphoadenylylsulfate(PAPS)-dependent sulfotransferase from arterial tissue ; it is specific for the N-and 0-sulfation of N-desulfoheparan sulfate. MATERIALS AND METHODS ChemicalsSephadex dextran gels and ConA-Sepharose were obtained from Pharmacia (Freiburg), DEAE-Trisacryl and Ultrogel AcA 202 from LKB (Uppsala), reference proteins for relative molecular mass determination from Serva (He...

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Section: Substratesmentioning

confidence: 99%

Section: Substratesmentioning

confidence: 99%

Purification and characterization of 3′‐phosphoadenylylsulfate: N‐desulfoheparan sulfate sulfotransferase from arterial tissue

Göhler

Niemann

Buddecke

1984

European Journal of Biochemistry

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“…A 48mg. sample of the faster electrophoretic fraction was dinitrophenylated by the procedure of Durant, Hendrickson & Montgomery (1962), except that when all the reagent had been added the solution was kept overnight at room temperature in the dark. After extraction with ether, the solution was dialysed, concentrated under reduced pressure to about 5ml.…”

Section: Naclmentioning

confidence: 99%

Protein-polysaccharides of pig laryngeal cartilage

Muir¹,

Jacobs²

1967

Biochemical Journal

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1. Protein-polysaccharides of chondroitin 4-sulphate were extracted with neutral calcium chloride from pig laryngeal cartilage that was not completely homogenized. The protein-polysaccharides were purified by precipitation with 9-aminoacridine. On zone electrophoresis in compressed glass fibre at pH7.2 it was separated into two fractions, although two distinct zones were not obtained. These fractions, which had already been shown to differ in their antigenic determinants, also differed considerably in amino acid composition, total protein, hexose and glucosamine contents. 2. The fraction of higher mobility contained approx. 2% of protein and only traces of glucosamine. Serine and glycine accounted for over half the total amino acid residues, but aromatic, basic and sulphur-containing amino acids were not detected. The weight-average molecular weight, determined by sedimentation, was 230000. 3. Assuming that there was the same sequence of neutral sugars at the linkage points as in PP-L fraction (protein-polysaccharide light fraction), the approximate molar ratio of hexose to serine suggested that most of the serine residues were linked to chondroitin sulphate chains. Support for this was derived from the agreement between the weight-average molecular weight of the chondroitin sulphate-peptide after proteolysis, and the chain weight calculated from its serine content. The chain weight based on the serine content of the fraction of higher electrophoretic mobility was approximately similar. 4. In contrast, the fraction of lower electrophoretic mobility resembled PP-L fraction in its amino acid composition, protein and glucosamine contents. The presence of glucosamine, together with the higher hexose content, suggested that this fraction contained some keratan sulphate. 5. The relatively low molecular weight of the fraction of higher mobility enabled it to be extracted without complete disintegration of the cartilage. The unlikelihood of its being produced by autolytic enzymes is discussed.

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“…Heparin.-A reinvestigation of the structure of heparin has been under-taken by Durant, Hendrickson & Montgomery (205) in which results of periodate oxidation of native and de-N-sulphated heparin were correlated with destruction of the specific sugar residues. The initial rapid reaction with periodate was followed by a slower oxidation complicated by the liberation of inorganic sulfate.…”

Section: �Ucopolysaccharidesmentioning

confidence: 99%

Carbohydrate Metabolism

Ashwell¹

1964

Annu. Rev. Biochem.

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This review is not designed as a comprehensive survey of the literature on carbohydrate metabolism, but has been restricted to five major areas of current activity. This restriction has permitted a more detailed consideration of the experimental data in order to provide a more critical basis for evalu ation of the significance of the work reported. NUCLEOTIDE SUGARSAs a result of intense interest during the last several years, a vast array of sugar nucleotides has been isolated and characterized. There has been, however, a marked shift of emphasis towards the study of nucleotide sugar dependent biosynthetic reactions involved in the formation of complex macromolecules. A striking example is seen in the development in knowledge of the structure of the bacterial O-antigenic polysaccharides, discussed in the section on lipopolysaccharides. Two excellent and comprehensive reviews covering the general area of sugar nucleotides and the synthesis of carbohy drates are currently in press (1, 2), and a complete survey of the biosynthetic reactions involved in the formation of deoxysugars has been published recently (3).Isolation and formation of sugar nucleotides.-Recondo & Leloir (4) found the rate of starch biosynthesis in plant tissues to be approximately tenfold greater when adenosine diphosphoglucose was substituted for uridine diphos phoglucose as precursor. This surprising observation prompted a search for supportive evidence that the former was a naturally occurring metabolite. Two such lines of evidence have now been adduced. Espada (5) isolated and partially purified (20-fold) an enzyme from wheat (ADP-glucose pyrophos phorylase) which catalyzes the formation of ADP-glucose and pyrophosphate upon incubation with ATP and glucose-I-phosphate. Resolution of this enzyme from UDP-glucose pyrophosphorylase by diethylaminoethyl cellu lose chromatography clearly established the separate nature of the two enzymes. The second piece of supporting evidence has been the identification of ADP-glucose as a naturally occurring compound (6) . From the alcoholic extract of one kilogram of corn grain, 8 JLmoles were obtained of a substance which behaved similarly to synthetic ADP-glucose upon paper chromatog raphy and electrophoresis. Upon mild acid hydrolysis, 60-70 percent of the 1 This review covers essentially the period from midsummer 1962 through August 1963.2 Among the abbreviations used in this review are: CMP, CDP, CTP (cytidine mono-, di-, and triphosphate); d (deoxy-); GDP (guanosine diphosphate); TDP (thymidine diphosphate); UDP (uridine diphosphate). 101 Annu. Rev. Biochem. 1964.33:101-138. Downloaded from www.annualreviews.org Access provided by University of California -San Francisco UCSF on 12/14/14. For personal use only. Quick links to online content Further ANNUAL REVIEWS

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Studies on the structure of heparin

Cited by 34 publications

References 24 publications

Purification and characterization of 3′‐phosphoadenylylsulfate: N‐desulfoheparan sulfate sulfotransferase from arterial tissue

Purification and characterization of 3′‐phosphoadenylylsulfate: N‐desulfoheparan sulfate sulfotransferase from arterial tissue

Protein-polysaccharides of pig laryngeal cartilage

Carbohydrate Metabolism

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