Heparin protein

Veldkamp CT, Peterson FC, Pelzek AJ, Volkman BF. The monomer-dimer equilibrium of stromal cell-derived factor-1 (CXCL 12) is altered by pH, phosphate, sulfate, and heparin. Protein Sci 2005 14 1071-81. 

Stuckey JA, St Charles R, Edwards BF. A model of the platelet factor 4 complex with heparin. Proteins 1992 14 277-87. 

Most of the common methods of isolation of heparin (described in sufficient detail in monographs128-30) are based on a procedure, developed by Charles and Scott,31 involving autolysis of the tissue (originally beef liver and beef lung), extraction with alkali, coagulation of proteins by heating, and precipitation of a heparin - protein complex by acidification. Heparin is recovered from the complex by reprecipitation with ethanol, or acetone, or both. Fats are removed by extraction with ethanol, and proteins by treatment with trypsin. Modifications of this proce-... 

Capila I, Linhardt RJ (2002) Heparin - Protein interactions. Angew Chem Int Ed 41 391—412. 

Mast Cells and Basophils. The chief sites of histamine storage are mast cells in the tissues and basophils in blood. These cells synthesize histamine and store it in secretory granules along with a heparin-protein complex. In response to specific antigens, mast cells or basophils are sensitized. Histamine is then secreted from the storage granules. Besides the histamine stores in mast cells and basophils, there is evidence of non-mast cell histamine in some tissues, particularly gastric and intestinal mucosa (60). 

Heparin protein interactions. In Lane DA et al (eds) Heparin and related polysaccharides. Plenum Press, New York, p 141... 

Table I. Antithrombinie and lytic properties of some heparin-protein complexes35 ...

The kinetics of the lytic effect displayed by the complexes of immobilized heparin with thrombin and fibrinogen, in distinction from those with plasmin, are described by their saturation curves. The observed slowing down of the dissolution of unstabilized fibrin is probably due to the inhibiton of the lytic activity of the complexes by the soluble products of the reaction. In fact, as it was shown in Ref. 106, further addition of immobilized heparin-protein complex to partially hydrolyzed fibrin results in a complete recovery of the dissolution rate. 

Capila I, Linhardt RJ Heparin-protein interactions. Angew Chemie Int. Ed. (2002)... 

Noti C, de Paz JL, Polito L, Seeberger PH. Preparation and use of microarrays containing synthetic heparin oligosaccharides for the rapid analysis of heparin-protein interactions. Chem. A Euro. J. 2006 12 8664-8686. 

Beef liver (or lung) was minced and then autolyzed for twenty-four hours before extraction with an alkaline solution saturated with ammonium sulfate. Protein was precipitated by warming the extract, and the heparin-protein complex was precipitated from the supernatant liquor on acidification. Extraction of the complex with ethanol removed fatty material, and tryptic digestion removed most of the protein. The heparin was precipitated with ethanol, redissolved in warm alkaline solution to destroy trypsin, and reprecipitated with acetone. This material, crude heparin, was isolated in a yield of 15-50 g. per 100 lb. of animal tissue. In a later paper," the purification of crude heparin by fractionation successively with Lloyd s reagent, cadmium chloride, and acetone, was described. The purified heparin w-as 100 times as active as the crude material. Scott and Charles" reported the presence of nitrogen... 

For the separation of heparin from heparin-protein complexes, Homan and Lens have developed a method which avoids the use of acid media. The heparin-protein complex is dissolved in aqueous solution at pH 7.5 and is extracted with phenol (which removes most of the protein). The method also facilitates the removal of colored impurities wLich are normally difficult to eliminate. Extraction of the heparin-protein-octylamine complex with phenol has been studied. "... 

Cardiogenic shock can occur in parallel with disseminated intravascular coagulation (5). In these circumstances heparin is thought to act as a hapten in a heparin-protein interaction that stimulates antibody production and an antigen-antibody reaction associated with release of platelet and vasoactive compounds. 

Capila I, Linhardt RJ (2002) Heparin-protein interactions. Angew Chem Int Ed 41 391 12 Carmeliet P (2003) Angiogenesis in health and disease. Nat Med 9 653-660 Chen CP, Imanishi Y, Ito Y (1998) Effect of protein and cell behavior on pattern-grafted thermoresponsive polymer. J Biomed Mater Res 42 38-44 Cheng XH, Canavan HE, Stein MJ et al (2005) Surface chemical and mechanical properties of plasma-polymerized N-isopropylacrylamide. Langmuir 21 7833-7841 Curti PS, de Moura MR, Veiga W et al (2005) Characterization of PNIPAAm photografted on PET and PS surfaces. Appl Surf Sci 245 223-233... 

L15. Lindahl, U., Further charcterisation of the heparin-protein linkage region. Biochim. Biophys. Acta 130, 368-382 (1966). 

L16. Lindahl, U., Structure of the heparin-protein linkage region. Isolation and characterisation of the disaccharide S-O- -n-glucuronosyl-n-galactose and the trisaccharide 3-0- -D-galactopvranosyl-4-0- 8-D-galactopyranosyl-D-xylose. Ark. Kemi 26, 101-110 (1967). 

Capila, I. and Linhardt, R. J. Heparin-protein interactions. Angew Chem Int Ed Engl, 41, 391, 2002. 

Capda, I., Linhardt, R. J. (2002). Heparin-protein interactions. Angewandte Chemie International Edition in English, 41(3), 391—412. Retrieved from, http //www.ncbi.nlm.nih. gov/entrez/query.fcgi cmd=Retrieve db=PubMed dopt=Citation list uids=12491369. 

It is advisable to have some ready-to-use commercially available affinity columns to hand, such as IMAC resins, p-aminobenzamidine. Reactive Blue, heparin. Protein A for antibody purification, as well as some activated resins such as tresyl, epoxy or N-hydroxysuccinimide. 

In view of the uncertainty as to the existence of a heparin-protein complex, Idndahl et al. (1965) analyzed a number of heparin samples for the presence of amino acids. It was found that heparin which had been prepared by use of mild methods, essentially involving only treatment with proteolytic enzymes, contained residual amino acids with serine as the main component. In some preparations serine was the only amino acid found in significant amounts. Samples prepared under more drastic conditions, including alkali extraction or bleaching, contained only traces of amino acids. These findings thus suggest that heparin does occur in the tissues as a covalently bound complex with protein. 

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