Effect of Deglycosylation on the Fibrin Polymerization Depending on NaCl Concentration

2013 ◽  
Vol 596 ◽  
pp. 213-218
Author(s):  
Kenji Kubota ◽  
Yoshiharu Toyama ◽  
Nobukazu Nameki ◽  
Kaori Wakamatsu
Keyword(s):  
Sialic Acid ◽  
Electrostatic Interactions ◽  
Carbohydrate Chain ◽  
Terminal Region ◽  
Fibrin Gel ◽  
Fibrin Polymerization ◽  
Amino Terminal ◽  
Terminal Sialic Acid ◽  
Nacl Concentration ◽  
Fibrinogen Molecule

Cleavage of carbohydrate chains linked to fibrinogen molecule results in an acceleration of fibrin polymerization, fibrin gel formation, by promoting the lateral aggregation of protofibrils. Sialic acid at the unreduced terminus of carbohydrate chain plays an essentially important role in the lateral aggre-gation. Fibrin polymerization is significantly affected by the solvent condition, e.g., pH and ionic strength. Terminal sialic acid are supposed to interact with amino terminal region of Bβ chain, in which there are many basic amino acid residues, and thus such interactions are expected to be electrostatic. In order to clarify whether the electrostatic interactions are essential for lateral aggregation, we examined temporal growth of fibrin polymerization of deglycosylated fibrinogen at high NaCl concentration. Marked acceleration of lateral aggregation was observed in deglycosylated fibrinogen even at high NaCl concentration where lateral aggregation was significantly inhibited in intact fibrinogen. These results suggest specific interactions of terminal sialic acid of the carbohydrate chain with the central E region of fibrinogen molecule, which may be important for the regulation of lateral aggregation rather than electrostatic interactions between the terminal sialic acids and the amino terminal region of Bβ chain.

Desialylation of N-Linked Carbohydrate Chain of Fibrinogen

2013 ◽  
Vol 534 ◽  
pp. 241-246
Author(s):  
Kenji Kubota ◽  
Yoshiharu Toyama ◽  
Nobukazu Nameki ◽  
Kaori Wakamatsu
Keyword(s):  
Sialic Acid ◽  
Carbohydrate Chain ◽  
Sialic Acids ◽  
Polymerization Process ◽  
Fibrin Polymerization ◽  
Terminal Sialic Acid ◽  
Mixing Effects ◽  
Polymerization Behavior ◽  
Carbohydrate Chains

Acceleration of fibrin polymerization occurs by the cleavage of sialic acids at the nonreducing terminal ends of N-linked carbohydrate chains as well as the cleavage of the entity of carbohydrate chains. In order to characterize and clarify the role of terminal sialic acid in the fibrin polymerization, mixing effects of desialylated fibrinogen with the intact one on the polymerization behavior were investigated by turbidity measurements in the course of polymerization. Marked accelerated fibrin polymerization was observed for the mixing of even a little amount of desialylated fibrinogen. Cleavage of the terminal sialic acid resulted in almost the equivalent accelerating effect to those of the deglycosylated fibrinogen, in which the entity of N-linked carbohydrate chain was cleaved. These results suggest that the terminal sialic acids regulate the fibrin polymerization in an inhibitory manner, and the cleavage of them induces the switchover from the protofibril growth to the lateral aggregation of fibrin polymerization process, resulting in the preferential fibrin polymerization.

scholarly journals Sialic acid prevents loss of large von Willebrand factor multimers by protecting against amino-terminal proteolytic cleavage

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1790-1796 ◽  
Author(s):  
SD Berkowitz ◽  
AB Federici
Keyword(s):  
Sialic Acid ◽  
Von Willebrand Factor ◽  
Proteinase Inhibitors ◽  
Terminal Region ◽  
Cleavage Sites ◽  
Amino Terminal ◽  
Carboxyl Terminal ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Beta Galactosidase

Abstract Removal of sialic acid from the von Willebrand factor (vWF) subunit exposes additional cleavage sites in the amino-terminal region that are associated with loss of large multimers. The extent of large multimer loss was evaluated by examining the sites of subunit cleavage of native and carbohydrate-modified vWF after treatment with trypsin, chymotrypsin, or plasmin. In the presence of proteinase inhibitors, purified vWF was treated with neuraminidase alone to remove 90% to 95% of the sialic acid or with neuraminidase and beta-galactosidase to remove the sialic acid and 45% to 50% of the D-galactose, with little or no loss of large multimers observed. Digestion of native vWF with trypsin produced the greatest loss of large multimers, while chymotrypsin produced less and plasmin produced the least. Large multimer loss was more extensive with each enzyme after carbohydrate modification of vWF. The extent and approximate location of subunit cleavage was determined by immunoblotting and monoclonal antibody epitope mapping. Trypsin, chymotrypsin, and plasmin were shown to produce both amino- and carboxyl-terminal fragments. The number, location, and relative quantities of carboxyl-terminal fragments produced were unchanged after carbohydrate modification. However, digestion of the amino-terminal region was considerably more extensive after carbohydrate modification as judged by a marked decrease or absence of the larger fragments seen when native vWF was digested, and by the appearance of new smaller molecular mass species. Therefore, the greater loss of large multimers that occurs after carbohydrate modification is likely to be the result of cleavages in the amino- terminal region of the molecule. By protecting the vWF subunit against amino-terminal cleavage, sialic acid inhibits the loss of large multimers.

scholarly journals Sialic acid prevents loss of large von Willebrand factor multimers by protecting against amino-terminal proteolytic cleavage

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1790-1796 ◽  
Author(s):  
SD Berkowitz ◽  
AB Federici
Keyword(s):  
Sialic Acid ◽  
Von Willebrand Factor ◽  
Proteinase Inhibitors ◽  
Terminal Region ◽  
Cleavage Sites ◽  
Amino Terminal ◽  
Carboxyl Terminal ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Beta Galactosidase

Removal of sialic acid from the von Willebrand factor (vWF) subunit exposes additional cleavage sites in the amino-terminal region that are associated with loss of large multimers. The extent of large multimer loss was evaluated by examining the sites of subunit cleavage of native and carbohydrate-modified vWF after treatment with trypsin, chymotrypsin, or plasmin. In the presence of proteinase inhibitors, purified vWF was treated with neuraminidase alone to remove 90% to 95% of the sialic acid or with neuraminidase and beta-galactosidase to remove the sialic acid and 45% to 50% of the D-galactose, with little or no loss of large multimers observed. Digestion of native vWF with trypsin produced the greatest loss of large multimers, while chymotrypsin produced less and plasmin produced the least. Large multimer loss was more extensive with each enzyme after carbohydrate modification of vWF. The extent and approximate location of subunit cleavage was determined by immunoblotting and monoclonal antibody epitope mapping. Trypsin, chymotrypsin, and plasmin were shown to produce both amino- and carboxyl-terminal fragments. The number, location, and relative quantities of carboxyl-terminal fragments produced were unchanged after carbohydrate modification. However, digestion of the amino-terminal region was considerably more extensive after carbohydrate modification as judged by a marked decrease or absence of the larger fragments seen when native vWF was digested, and by the appearance of new smaller molecular mass species. Therefore, the greater loss of large multimers that occurs after carbohydrate modification is likely to be the result of cleavages in the amino- terminal region of the molecule. By protecting the vWF subunit against amino-terminal cleavage, sialic acid inhibits the loss of large multimers.

scholarly journals SEL-5, A Serine/Threonine Kinase That Facilitates lin-12 Activity in Caenorhabditis elegans

Genetics ◽  
1999 ◽  
Vol 153 (4) ◽  
pp. 1641-1654 ◽  
Author(s):  
Hanna Fares ◽  
Iva Greenwald
Keyword(s):  
Cell Fate ◽  
Point Mutations ◽  
Kinase Domain ◽  
Terminal Region ◽  
Serine Threonine Kinase ◽  
Intracellular Domain ◽  
Cell Fate Decisions ◽  
Threonine Kinase ◽  
Amino Terminal ◽  
Fate Decision

Abstract Ligands present on neighboring cells activate receptors of the LIN-12/Notch family by inducing a proteolytic cleavage event that releases the intracellular domain. Mutations that appear to eliminate sel-5 activity are able to suppress constitutive activity of lin-12(d) mutations that are point mutations in the extracellular domain of LIN-12, but cannot suppress lin-12(intra), the untethered intracellular domain. These results suggest that sel-5 acts prior to or during ligand-dependent release of the intracellular domain. In addition, sel-5 suppression of lin-12(d) mutations is tissue specific: loss of sel-5 activity can suppress defects in the anchor cell/ventral uterine precursor cell fate decision and a sex myoblast/coelomocyte decision, but cannot suppress defects in two different ventral hypodermal cell fate decisions in hermaphrodites and males. sel-5 encodes at least two proteins, from alternatively spliced mRNAs, that share an amino-terminal region and differ in the carboxy-terminal region. The amino-terminal region contains the hallmarks of a serine/threonine kinase domain, which is most similar to mammalian GAK1 and yeast Pak1p.

scholarly journals Role of the amino-terminal region of the DnaA protein in opening of the duplex DNA at theoriCregion

1999 ◽  
Vol 176 (1) ◽  
pp. 163-167 ◽  
Author(s):  
Shinji Mima ◽  
Yoshihiro Yamagachi ◽  
Taemi Kondo ◽  
Tomofusa Tsuchiya ◽  
Tohru Mizushima
Keyword(s):  
Terminal Region ◽  
Duplex Dna ◽  
Dnaa Protein ◽  
Amino Terminal

scholarly journals Molecular cloning of the amino-terminal region of a rat MUC 2 mucin gene homologue. Evidence for expression in both intestine and airway.

1994 ◽  
Vol 269 (27) ◽  
pp. 17833-17840
Author(s):  
H. Ohmori ◽  
A.F. Dohrman ◽  
M. Gallup ◽  
T. Tsuda ◽  
H. Kai ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Terminal Region ◽  
Amino Terminal ◽  
Mucin Gene ◽  
Gene Homologue

scholarly journals Characterization of Virus Adsorption by Using DEAE-Sepharose and Octyl-Sepharose

2002 ◽  
Vol 68 (8) ◽  
pp. 3965-3968 ◽  
Author(s):  
Patricia A. Shields ◽  
Samuel R. Farrah
Keyword(s):  
Sodium Chloride ◽  
Electrostatic Interactions ◽  
Hydrophobic Interactions ◽  
Chloride Concentration ◽  
Sodium Chloride Concentration ◽  
Virus Adsorption ◽  
Nacl Concentration

ABSTRACT Viruses were characterized by their adsorption to DEAE-Sepharose or by their elution from octyl-Sepharose by using buffered solutions of sodium chloride with different ionic strengths. Viruses whose adsorption to DEAE-Sepharose was reduced most rapidly by an increase in the sodium chloride concentration were considered to have the weakest electrostatic interactions with the solids; these viruses included MS2, E1, and φX174. Viruses whose adsorption to DEAE-Sepharose was reduced least rapidly were considered to have the strongest electrostatic interactions with the column; these viruses included P1, T4, T2, and E5. All of the viruses studied adsorbed to octyl-Sepharose in the presence of 4 M NaCl. Viruses that were eluted most rapidly following a decrease in the concentration of NaCl were considered to have the weakest hydrophobic interactions with the column; these viruses included φX174, CB4, and E1. Viruses that were eluted least rapidly from the columns after the NaCl concentration was decreased were considered to have the strongest hydrophobic interactions with the column; these viruses included f2, MS2, and E5.

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