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

The arrangement of disulfide bonds in fragment D from human fibrinogen

1978 ◽  
Vol 13 (3) ◽  
pp. 557-562 ◽  
Author(s):  
Hessel Bouma ◽  
Takashi Takagi ◽  
Russell F. Doolittle
Keyword(s):  
Disulfide Bonds ◽  
Human Fibrinogen

scholarly journals Amino Acid Sequence Studies on the β-Chain of Human Fibrinogen

1977 ◽  
Author(s):  
K. Watt ◽  
D. Goldbaum ◽  
B. A. Cottrell ◽  
T. Takagi ◽  
R. F. Doolittle
Keyword(s):  
Disulfide Bonds ◽  
Coiled Coil ◽  
Enzymatic Digestion ◽  
The Other ◽  
Human Fibrinogen ◽  
Parent Molecule ◽  
General Arrangement ◽  
The One ◽  
Β Chain

The β-chain of human fibrinogen contains 480 ± 15 residues, sixteen of which are methionines. In this vein, we have isolated and characterized all seventeen cyanogen bromide peptides. The arrangement of most of these fragments has been achieved by the identification of key overlap peptides derived from enzymatic digestion of β-chains, on the one hand, and a characterization of β-chain fragments isolated from fragments D and E, on the other. In some cases the alignment is based only on homologies with the α- and/or γ-chains, and in a few instances some definite ambiguities still exist. For the most part, however, the general arrangement is in hand. Of particular interest is the plasmin-sensitive segment which is a part of the inter-domainal connection, which in turn we believe is a three-stranded coiled-coil punctuated by an unusual arrangement of disulfide bonds. The positioning of the other cysteine residues in the β-chain is also of considerable interest, since it sheds light on the extent of connections between portions of the three non-identical chains in the parent molecule.

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 Amino Acid Sequence Studies on Human Fibrinogen:Arrangement of Interchain Disulfide Bonds Bounding the Regions of Three-Stranded Ropes

1977 ◽  
Author(s):  
R. F. Doolittle ◽  
D. Goldbaum ◽  
L. R. Doolittle
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Disulfide Bonds ◽  
Molecular Model ◽  
Coiled Coils ◽  
Human Fibrinogen ◽  
Type Molecule ◽  
Significant Homology ◽  
Atomic Coordinates

Human fibrinogen contains 29 disulfide bonds, only three of which are involved in holding the two dimeric halves of the molecule together. Of the remainder, twelve others are arranged in four sets of three bonds each. Thus, each half of the molecule has two of these unusual arrangements separated by a three-stranded rope consisting of approximately 110 residues in each chain. Alignment of the three non-identical chains at the appropriate cysteine residues participating in these assemblies has revealed not only significant homology but also a rhythmic occurrence of polar and nonpolar amino acids consistent with the existence of coiled α-helices. Indeed, other workers had predicated the existence of such coiled-coils on the basis of fiber diffraction studies a generation ago, hypothesizing that they were likely inter-domainal connections holding the distal portions of a Hall and Slayter-type molecule to the central region. Our data indicate that these predictions were essentially correct. To reinforce the point we have constructed a detailed molecular model of the connecting regions, even to the point where the hypothetical atomic coordinates have been recorded. The model is consistent with virtually all physical data and illuminates details of how molecular packing may occur during formation. It also delineates the geography of bond splitting during fibrinolysis, especially with regard to the boundaries of fragments D and E.

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.

Cryo-electron microscopy of two-dimensional crystals of reduced type B botulinum neurotoxin

1992 ◽  
Vol 50 (1) ◽  
pp. 530-531
Author(s):  
P. F. Flicker ◽  
V.S. Kulkarni ◽  
J. P. Robinson ◽  
G. Stubbs ◽  
B. R. DasGupta
Keyword(s):  
Disulfide Bonds ◽  
Clostridium Botulinum ◽  
Structural Changes ◽  
Two Dimensional ◽  
Type B ◽  
Single Chain ◽  
Muscle Paralysis ◽  
Cryo Electron Microscopy ◽  
Disulfide Linkages ◽  
Intracellular Action

Botulinum toxin is a potent neurotoxin produced by Clostridium botulinum. The toxin inhibits release of neurotransmitter, causing muscle paralysis. There are several serotypes, A to G, all of molecular weight about 150,000. The protein exists as a single chain or or as two chains, with two disulfide linkages. In a recent investigation on intracellular action of neurotoxins it was reported that type B neurotoxin can inhibit the release of Ca++-activated [3H] norepinephrine only if the disulfide bonds are reduced. In order to investigate possible structural changes in the toxin upon reduction of the disulfide bonds, we have prepared two-dimensional crystals of reduced type B neurotoxin. These two-dimensional crystals will be compared with those of the native (unreduced) type B toxin.

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