Structural Studies of Heparin. I. Hydrolytic Cleavage of Sulfates<sup>*</sup>

Helbert, J.; Marini, Mario

doi:10.1021/bi00905a033

Cited by 47 publications

(5 citation statements)

References 15 publications

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“…The plasmon resonance studies also permit an estimate of the dissociation constant of aFGF for heparin, which is found to be on the order of 50-140 nM and is in excellent agreement with that obtained from affinity electrophoresis experiments (91 nM; Lee & Lander, 1991). The chemical modifications used in these experiments alter different functional groups of heparin to varying extents (Helbert & Marini, 1963;Chung & Ellerton, 1976), so it is not surprising that a range of Xd, kon, and k0iT values was observed on the two surfaces.…”

Section: Discussionsupporting

confidence: 74%

Nature of the interaction of heparin with acidic fibroblast growth factor

Mach¹,

Volkin²,

Burke³

et al. 1993

Biochemistry

194

130

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The binding of human acidic fibroblast growth factor (aFGF) to heparin has been analyzed by a variety of different approaches to better elucidate the nature of this protein/sulfated polysaccharide interaction. Static and dynamic light scattering as well as analytical ultracentrifugation analyses indicates that 14-15 molecules of a FGF can bind to a 16-kDa heparin chain, with approximately 10 of these bound relatively uniformly to high-affinity sites. The dissociation constants of these latter sites are estimated to be approximately 50-140 nM on the basis of surface plasmon resonance experiments in which the association and dissociation rates of aFGF interaction with immobilized heparin were measured. The size of the binding site of a FGF on heparin was also determined by heparin lyase digestion of a FGF/heparin complexes followed by isolation and characterization of protected oligosaccharides. The smallest aFGF-protected oligosaccharide comigrated with delta UA2S(1-->4)-alpha-D-GlcNp2S6S(1-->4)-alpha-L-IdoAp-2S( 1-->4)-alpha-D-GlcNp2S6S (where delta UA represents 4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid and S is sulfate). Thus, aFGF appears to bind at high density (one molecule every 4-5 polysaccharide units) and with high affinity to heparin. This potentially provides a concentrated, stabilized storage form of the growth factor that can be released for receptor-mediated cellular activation in response to the proper stimuli. It is also possible that close proximity of aFGF molecules on the highly sulfated regions of heparan chains may be involved in the induction of receptor aggregation as suggested by Ornitz et al. [Ornitz, D. M., Yayon, A., Flanagan, J. G., Svahn, C. M., Levi, E., & Leder, P. (1992) Mol. Cell. Biol. 12, 240-247].

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Section: Discussionsupporting

confidence: 74%

Nature of the interaction of heparin with acidic fibroblast growth factor

Mach¹,

Volkin²,

Burke³

et al. 1993

Biochemistry

194

130

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“…Mucopolysac- well known, 11-14 with that from shark (S-Ch-68, not shown) having a greater sulfate/ sugar nitrogen ratio (up to 1.6). 16 The structure for heparin was taken from recent ~t u d i e s~~J~ with the broken line indicating that the sulfate groups, although largely at the 6 position of the amino sugar, may occur elsewhere in the dimeric unit. The structure of a variety of kersto sulfates is still under investigation18-20 although the well-known form for KS-121*22 is shown in the figure.…”

Section: Resultsmentioning

confidence: 99%

Optical rotatory dispersion of mucopolysaccharides and mucopolysaccharide–dye complexes. II. Ultraviolet cotton effects in the amide transition bands

Stone

1969

Biopolymers

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synopsisUltraviolet optical rotatory dispersion curves of mucopolysaccharides exhibit particular Cotton effects in the spectral region of the It-x and T-T amide transitions. Two general patterns emerge: (I) enhancement of negative rotation and of the first negative Cotton effect (troughs 217-220 mp) and (2) relative dominance of the positive Cotton effect in the 7r-if transition region (peak -198 mp). Groups (1) and (2) can be correlated with a structural difference in the linkages of the amino sugars: (1) occurs with polymers containing 3-1-linked glycosamino sugars and (2) with glycosamino moieties linked 4-1 by either a-or 8-glycosidic bonds Measurements of the circular dichroic absorption bands support the qualitative conclusions from optical rotation. AU mucopolysaccharides exhibit a first, negative band centered at 208-211 mp, while only those in group (2) show, in addition, a positive band centered at 189-192 mp. A suggested unifying model considers that difference in kind and/or degree of preferred geometry of the amide groups obtains from two forms of secondary order: (I) having a linear hydrogen bond from the N (acceptor) to the (C2)G-H of the preceding uronic acid and (2) having a linear hydrogen bond from the N (acceptor) to the (CZ or 8) (fH of the following sugar. The hydrogen bonds would have similar strength but opposite directions in the two systems [towards the nonreducing end (1) or towards the reducing end (2)], closing eight-membered rings

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“…The sodium heparinate which was obtained at the end of the purification process was designed as HEP Partially N-desulfated heparins of different degree of sulfation (X NDHEP with X = percentage of N-desulfation) were prepared by acid catalyzed cleavage of the glucosamine N-sulfamino groups of heparinic acid in aqueous solution as recommended by Helbert and Marini [15]. Total N-desulfation was achieved according to the method proposed by Inoue and Nagasawa [16].…”

Section: Resultsmentioning

confidence: 99%

Chemical Modifications of Heparin.1: Reaction of Partially N-Desulfated Heparin with Glutaraldehyde

Diancourt

Braud

Vert

1994

Journal of Bioactive and Compatible Polymers

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Purified glutaraldehyde was allowed to react with the primary amino groups generated by mild acidolysis of glucosamine N -sulfamino residues present in heparinic acid. The reaction yielded high molecular mass heparinoids (HMMHEP) or hydrogels, depending on experimental conditions. It was shown that a selected HMMHEP exhibited a higher anticoagulant activity than the starting heparin or the parent partially N -desulfated heparin. This HMMHEP degraded slowly in vitro in an aqueous medium buffered at pH = 7.4. Moreover, HMMHEP was biologically active after i.v. injection but not after s.c. and i.m. injections. Hydrogels also degraded slowly to yield biologically active higher molecular mass heparinoid. These findings suggested that neither N -desulfation nor coupling of glutaraldehyde adducts affected in a large extend the activity of the parent heparin.

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Structural Studies of Heparin. I. Hydrolytic Cleavage of Sulfates^*

Cited by 47 publications

References 15 publications

Nature of the interaction of heparin with acidic fibroblast growth factor

Nature of the interaction of heparin with acidic fibroblast growth factor

Optical rotatory dispersion of mucopolysaccharides and mucopolysaccharide–dye complexes. II. Ultraviolet cotton effects in the amide transition bands

Chemical Modifications of Heparin.1: Reaction of Partially N-Desulfated Heparin with Glutaraldehyde

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Structural Studies of Heparin. I. Hydrolytic Cleavage of Sulfates*

Cited by 47 publications

References 15 publications

Nature of the interaction of heparin with acidic fibroblast growth factor

Nature of the interaction of heparin with acidic fibroblast growth factor

Optical rotatory dispersion of mucopolysaccharides and mucopolysaccharide–dye complexes. II. Ultraviolet cotton effects in the amide transition bands

Chemical Modifications of Heparin.1: Reaction of Partially N-Desulfated Heparin with Glutaraldehyde

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Structural Studies of Heparin. I. Hydrolytic Cleavage of Sulfates^*