Formation of the Human Fibrinogen Subclass Fib420: Disulfide Bonds and Glycosylation in Its Unique (EChain) Domains

Blood ◽  
1998 ◽  
Vol 92 (9) ◽  
pp. 3302-3308 ◽  
Author(s):  
Yiping Fu ◽  
Jian-Zhong Zhang ◽  
Colvin M. Redman ◽  
Gerd Grieninger
Keyword(s):  
Disulfide Bonds ◽  
Cell Transfection ◽  
The Novel ◽  
Disulfide Bridges ◽  
Human Fibrinogen ◽  
Cos Cells ◽  
C Terminus ◽  
Symmetrical Molecule ◽  
American Society

Abstract COS cell transfection has been used to monitor the assembly and secretion of fibrinogen molecules, both those of the subclass containing the novel E chain and those of the more abundant subclass whose  chains lack E’s globular C-terminus. That region, referred to as the EC domain, is closely related to the ends of β and γ chains of fibrinogen (βC and γC). Transfection of COS cells with E, β, and γ cDNAs alone results in secretion of the symmetrical molecule (Eβγ)2, also known as Fib420. Cotransfection with cDNA for the shorter  chain yielded secretion of both (βγ)2 and (Eβγ)2 but no mixed molecules of the structure E(βγ)2. Exploiting the COS cells’ fidelity with regard to Fib420 production, identification was made of the highly conserved Asn667 as the sole site of N-linked glycosylation in the E chain. No evidence from Cys → Ser replacements was found for interchain disulfide bridges involving the four cysteines of the EC domain. However, for fibrinogen secretion, the E, β, and γ subunits do exhibit different requirements for integrity of the two intradomain disulfide bridges located at homologous positions in their respective C-termini, indicating dissimilar structural roles in the process of fibrinogen assembly. © 1998 by The American Society of Hematology.

Formation of the Human Fibrinogen Subclass Fib420: Disulfide Bonds and Glycosylation in Its Unique (EChain) Domains

Blood ◽  
1998 ◽  
Vol 92 (9) ◽  
pp. 3302-3308 ◽  
Author(s):  
Yiping Fu ◽  
Jian-Zhong Zhang ◽  
Colvin M. Redman ◽  
Gerd Grieninger
Keyword(s):  
Disulfide Bonds ◽  
Cell Transfection ◽  
The Novel ◽  
Disulfide Bridges ◽  
Human Fibrinogen ◽  
Cos Cells ◽  
C Terminus ◽  
Symmetrical Molecule ◽  
American Society

COS cell transfection has been used to monitor the assembly and secretion of fibrinogen molecules, both those of the subclass containing the novel E chain and those of the more abundant subclass whose  chains lack E’s globular C-terminus. That region, referred to as the EC domain, is closely related to the ends of β and γ chains of fibrinogen (βC and γC). Transfection of COS cells with E, β, and γ cDNAs alone results in secretion of the symmetrical molecule (Eβγ)2, also known as Fib420. Cotransfection with cDNA for the shorter  chain yielded secretion of both (βγ)2 and (Eβγ)2 but no mixed molecules of the structure E(βγ)2. Exploiting the COS cells’ fidelity with regard to Fib420 production, identification was made of the highly conserved Asn667 as the sole site of N-linked glycosylation in the E chain. No evidence from Cys → Ser replacements was found for interchain disulfide bridges involving the four cysteines of the EC domain. However, for fibrinogen secretion, the E, β, and γ subunits do exhibit different requirements for integrity of the two intradomain disulfide bridges located at homologous positions in their respective C-termini, indicating dissimilar structural roles in the process of fibrinogen assembly. © 1998 by The American Society of Hematology.

scholarly journals Fib420 , the Novel Fibrinogen Subclass: Newborn Levels Are Higher Than Adult

Blood ◽  
1997 ◽  
Vol 90 (7) ◽  
pp. 2609-2614 ◽  
Author(s):  
Gerd Grieninger ◽  
Xianghan Lu ◽  
Yan Cao ◽  
Yiping Fu ◽  
Bohdan J. Kudryk ◽  
...  
Keyword(s):  
Developmental Expression ◽  
Striking Difference ◽  
The Novel ◽  
Western Blot Assay ◽  
Human Fibrinogen ◽  
C Terminus ◽  
Wide Range ◽  
Significant Difference ◽  
Adult Plasma ◽  
Normal Human

Abstract Fib420 is a recently identified subclass of normal human fibrinogen in which two extended α chain isoforms (αE ) replace the common α chains, yielding a molecule (ca. 420 kD) which is larger than the more abundant 340-kD form. Evidence for preservation of this subclass throughout vertebrate evolution suggests it performs some as yet unidentified vital function. A survey was undertaken to establish the range of plasma Fib420 levels in normal, healthy adults and in placental cord (fetal) blood. For measuring Fib420 , a quantitative Western blot assay was developed using monoclonal antibody against the exon-VI encoded C-terminus of the molecule's unique αE chain. This αE chain signal was normalized to that of the β chain, common to both fibrinogen forms. Analysis of plasma samples from the adult and newborn cohorts (n = 25 each; total fibrinogen ca. 2.6 mg/mL in both) revealed a statistically significant difference, with a mean level of 100 ± 28 μg/mL in the neonate compared to 34 ± 7 μg/mL in the adult. On average, 1 out of every 100 fibrinogen molecules in adult plasma belongs to the Fib420 subclass. Unlike in the newborn, adult Fib420 levels remained the same over a wide range of total plasma fibrinogen. The striking difference observed between these two cohorts suggests a changing developmental expression of the Fib420 subclass and a homeostatic control operating in later stages of life.

scholarly journals Fib420 , the Novel Fibrinogen Subclass: Newborn Levels Are Higher Than Adult

Blood ◽  
1997 ◽  
Vol 90 (7) ◽  
pp. 2609-2614 ◽  
Author(s):  
Gerd Grieninger ◽  
Xianghan Lu ◽  
Yan Cao ◽  
Yiping Fu ◽  
Bohdan J. Kudryk ◽  
...  
Keyword(s):  
Developmental Expression ◽  
Striking Difference ◽  
The Novel ◽  
Western Blot Assay ◽  
Human Fibrinogen ◽  
C Terminus ◽  
Wide Range ◽  
Significant Difference ◽  
Adult Plasma ◽  
Normal Human

Fib420 is a recently identified subclass of normal human fibrinogen in which two extended α chain isoforms (αE ) replace the common α chains, yielding a molecule (ca. 420 kD) which is larger than the more abundant 340-kD form. Evidence for preservation of this subclass throughout vertebrate evolution suggests it performs some as yet unidentified vital function. A survey was undertaken to establish the range of plasma Fib420 levels in normal, healthy adults and in placental cord (fetal) blood. For measuring Fib420 , a quantitative Western blot assay was developed using monoclonal antibody against the exon-VI encoded C-terminus of the molecule's unique αE chain. This αE chain signal was normalized to that of the β chain, common to both fibrinogen forms. Analysis of plasma samples from the adult and newborn cohorts (n = 25 each; total fibrinogen ca. 2.6 mg/mL in both) revealed a statistically significant difference, with a mean level of 100 ± 28 μg/mL in the neonate compared to 34 ± 7 μg/mL in the adult. On average, 1 out of every 100 fibrinogen molecules in adult plasma belongs to the Fib420 subclass. Unlike in the newborn, adult Fib420 levels remained the same over a wide range of total plasma fibrinogen. The striking difference observed between these two cohorts suggests a changing developmental expression of the Fib420 subclass and a homeostatic control operating in later stages of life.

Dimeric half-molecules of human fibrinogen are joined through disulfide bonds in an antiparallel orientation

Biochemistry ◽  
1983 ◽  
Vol 22 (9) ◽  
pp. 2049-2055 ◽  
Author(s):  
Paul D. Hoeprich ◽  
Russell F. Doolittle
Keyword(s):  
Disulfide Bonds ◽  
Human Fibrinogen

Thermal Conductivity Performance Modeling and Characterization of a Novel Anisotropic Conductive Adhesive Used in Lead-Free Electronics Packaging

2012 ◽  
Author(s):  
Matthew J. Combs ◽  
S. Manian Ramkumar ◽  
Satish Kandlikar
Keyword(s):  
Thermal Conductivity ◽  
Base Material ◽  
Bond Line ◽  
Thermal Interface Material ◽  
The Novel ◽  
Conductive Adhesives ◽  
Curing Conditions ◽  
Bond Line Thickness ◽  
Significant Research ◽  
American Society

The continued desire to utilize an alternative to lead-based solder materials for electrical interconnections has led to significant research interest in Anisotropic Conductive Adhesives (ACAs). The use of ACAs in electrical connections creates bonds using a combination of metal particles and epoxies to replace solder. The novel ACA discussed in this paper allows for bonds to be created through aligning columns of conductive particles along the Z-axis. These columns are formed by the application of a magnetic field, during the curing process. The benefit of this novel ACA is that it does not require precise printing of the adhesive on pads and also enables the mass curing without creating shorts in the circuitry. This paper will present the findings of the thermal conductivity performance tests using the novel ACA and its applicability as a thermal interface material and for assembling bottom termination components, power devices, etc. The columns that act as electrical conduction paths also contribute towards the thermal conductivity. The thermal conductivity of the novel ACA was measured utilizing a system that is similar to that in ASTM (American Society of Testing Materials) D5470 standard. The goal was to examine the influence of Bond Line Thickness (BLT), particle loading densities, particle diameters and adhesive matrix curing conditions on the electrical and thermal performance of the novel ACA. This paper will also present a numerical model to describe the thermal behavior of the novel ACA. The novel ACA’s applicability for PCB-level assembly has also been successfully demonstrated by RIT, including base material characterization, effect of process parameters, failures, and long-term reliability. Reliability testing included the investigation of the assembly performance in temperature and humidity aging, thermal aging, air-to-air thermal cycling, and drop testing.

scholarly journals Stabilization of phage T4 lysozyme by engineered disulfide bonds

1989 ◽  
Vol 86 (17) ◽  
pp. 6562-6566 ◽  
Author(s):  
M Matsumura ◽  
W J Becktel ◽  
M Levitt ◽  
B W Matthews
Keyword(s):  
Disulfide Bonds ◽  
Theoretical Calculations ◽  
Equilibrium Constants ◽  
T4 Lysozyme ◽  
Disulfide Bridges ◽  
Large Loop ◽  
Phage T4 ◽  
Flexible Part ◽  
Free Phage

Four different disulfide bridges (linking positions 9-164, 21-142, 90-122, and 127-154) were introduced into a cysteine-free phage T4 lysozyme at sites suggested by theoretical calculations and computer modeling. The new cysteines spontaneously formed disulfide bonds on exposure to air in vitro. In all cases the oxidized (crosslinked) lysozyme was more stable than the corresponding reduced (noncrosslinked) enzyme toward thermal denaturation. Relative to wild-type lysozyme, the melting temperatures of the 9-164 and 21-142 disulfide mutants were increased by 6.4 degrees C and 11.0 degrees C, whereas the other two mutants were either less stable or equally stable. Measurement of the equilibrium constants for the reduction of the engineered disulfide bonds by dithiothreitol indicates that the less thermostable mutants tend to have a less favorable crosslink in the native structure. The two disulfide bridges that are most effective in increasing the stability of T4 lysozyme have, in common, a large loop size and a location that includes a flexible part of the molecule. The results suggest that stabilization due to the effect of the crosslink on the entropy of the unfolded polypeptide is offset by the strain energy associated with formation of the disulfide bond in the folded protein. The design of disulfide bridges is discussed in terms of protein flexibility.

Fibrinogen is required for maintenance of platelet intracellular and cell-surface P-selectin expression

Blood ◽  
2009 ◽  
Vol 114 (2) ◽  
pp. 425-436 ◽  
Author(s):  
Hong Yang ◽  
Sean Lang ◽  
Zhimin Zhai ◽  
Ling Li ◽  
Walter H. A. Kahr ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Surface ◽  
Human Fibrinogen ◽  
C Terminus ◽  
Β3 Integrin ◽  
Von Willebrand ◽  
Selectin Binding ◽  
Willebrand Factor ◽  
Deficient Mice ◽  
Granule Release

Abstract Platelet P-selectin plays important roles in inflammation and contributes to thrombosis and hemostasis. Although it has been reported that von Willebrand factor (VWF) affects P-selectin expression on endothelial cells, little information is available regarding regulation of platelet P-selectin expression. Here, we first observed that P-selectin expression was significantly decreased on platelets of fibrinogen and VWF double-deficient mice. Subsequently, we identified this was due to fibrinogen deficiency. Impaired P-selectin expression on fibrinogen-deficient platelets was further confirmed in human hypofibrinogenemic patients. We demonstrated that this impairment is unlikely due to excessive P-selectin shedding, deficient fibrinogen-mediated cell surface P-selectin binding, or impaired platelet granule release, but rather is due to decreased platelet P-selectin content. Fibrinogen transfusion completely recovered this impairment in fibrinogen-deficient (Fg−/−) mice, and engagement of the C-terminus of the fibrinogen γ chain with β3 integrin was required for this process. Furthermore, Fg−/− platelets significantly increased P-selectin expression following transfusion into β3 integrin–deficient mice and when cultured with fibrinogen. These data suggest fibrinogen may play important roles in inflammation, thrombosis, and hemostasis via enhancement of platelet P-selectin expression. Since human fibrinogen levels vary significantly in normal and diseased populations, P-selectin as an activation marker on platelets should be used with caution.

scholarly journals Repercussions of Stereotyping and Cultural Bigotry in John Updike’s 9/11 Fiction

2021 ◽  
Vol 3 (1) ◽  
pp. 64-73
Author(s):  
Saleem Dhobi
Keyword(s):  
African Americans ◽  
Ethnic Groups ◽  
Social Exclusion ◽  
Western Culture ◽  
Cultural Integration ◽  
The Novel ◽  
Cultural Groups ◽  
Religious Minorities ◽  
The Us ◽  
American Society

This article analyzes Updike’s 9/11 novel, Terrorist to explore the implications of stereotyping and cultural bigotry in US society in the aftermath. The novelist demonstrates the problematic in the cultural integration of minorities particularly Muslims and Jews as represented by Ahmad and Jack Levy. The primary motto of the article is to analyze the novel from the perspective of the protagonists Ahmad and Jack who suffer the cultural and social exclusion in American society. Ahmad is the victim of cultural bigotry and Jack Levy faces discriminatory practices at school. The isolation and marginalization of Ahmad and Jack respectively imply the ethnic crevices prevalent in the US society. The author demonstrates that the dominant cultural groups: European and African Americans do not accept the religious minorities: Muslims and Jews. Consequently, Muslims who are overtly the targets of cultural hatred and marginalization in the aftermath of the 9/11 as portrayed in the novel become hostile toward the Western culture. The efforts for integration of religious minorities are cosmetic as exemplified in the cases of Ahmad and Jack in the text. The writer makes a balance in representing both dominant and Muslim cultures to demonstrate the problems pertaining to ethnic groups at their failure in accommodating differences. The cultural separation and hatred prevalent in US society become obstacles even for those like Jack who seek to integrate. The paper eventually demonstrates the possibility of integration of religious minorities when both mainstream Americans and people of religious minorities conform to accepting the differences.

scholarly journals STATE OF EXCEPTION THROUGH RASISME IN GO SET A WATCHMAN IN AGAMBEN’S POLITICAL PHILOSOPHY

2021 ◽  
Vol 20 (2) ◽  
pp. 123-134
Author(s):  
Damay Rahmawati ◽  
Ramadhani Ardianto Karsa Sunaryono ◽  
Mira Utami
Keyword(s):  
Political Philosophy ◽  
Racial Discrimination ◽  
Black People ◽  
Lower Class ◽  
State Of Exception ◽  
The Novel ◽  
Citizenship Rights ◽  
American Society ◽  
The Impact ◽  
Harper Lee

This study aims to see racism in the novel Go Set a Watchman by Harper Lee as state of exception; a political philosophy of Agamben. Agamben's idea of ​​state of exception is used in this study as the theoretical framework. This research specifically reveals how racism becomes part of state of exception in American society around 1960s when the novel was written. The analysis focuses on issues of racism in American society as depicted in the novel. The issue of racism is taken with the aim of analyzing state of exception in USA, in dealing with racial discrimination. After analyzing the issues of racism and state of exception in the novel, this study reveals that racism in American society is politically structured. The finding of this study is the discrimination experienced by lower class citizens who are dominated by black people, as the impact of state of exception which affects their citizenship rights.

scholarly journals Self-Abdication and Otherness in Philip Roth’s The Human Stain (2000)

2019 ◽  
Author(s):  
Assia Mohdeb ◽  
Sofiane Mammeri
Keyword(s):  
Racial Identity ◽  
Identity Formation ◽  
Well Being ◽  
The Novel ◽  
Racial Passing ◽  
The Core ◽  
Class Discrimination ◽  
The Subject ◽  
American Society ◽  
Definition Of

Identity, in one of its understanding, signifies a set of characteristics that make up a person’s ethical faithfulness to, identification with, and pride of one’s origin, tradition, and culture. Remaining true to one’s identity and being faithful to the core values of one’s culture is a complicated matter when it comes to a black living in white society like America, where color and racial identity are rudimentary prerequisites in self-definition and naming. Philip Roth’s novel entitled The Human Stain (2000) shows how some black figures undress their black identity to wear the prestigious white one to go onward with life as full selves, to have access to all the privileges the whites enjoy, and, above all, to live without the specter of race and the decisiveness of epidermal signs. The novel calls into question and revision such essentialist notions as other, class,andrace by describing the crises the subject or self undergoes in the light of racial prejudices, center-periphery relations, and class stereotypes. The present paper, then, addresses the act of self-abdication the protagonist, Silk Coleman, carries out to overstep the feeling of otherness and to dodge racial discrimination. The paper looks into the notions of selfhood and Otherness by negotiating the definition of the self and the distortion it undergoes in its encounter with the Other . The study aims at revealing, primarily, the effects of Black racial-passing, a common phenomenon in American society of the first half of the twentieth century, on familial relationships and cultural heritage. It also reveals the weight of gender and class discrimination in the individual’s identity formation and well-being.

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