Fibrinogen Biosynthesis

A bstract : Plasma fibrinogen is synthesized primarily in hepatocytes and assembly of the three component chains (Aα, Bβ, and γ) into its final form as a six‐chain dimer (Aα, Bβ, γ) 2 occurs rapidly in the lumen of the endoplasmic reticulum (ER). Assembly takes place in a stepwise manner with single chains interacting with each other to form Aα‐γ and Bβ‐γ complexes. The two‐chain complexes then acquire another chain to form half‐molecules (Aα, Bβ, γ) 1 , which in a final step are linked to form the six‐chain (Aα, Bβ, γ) 2 complex. As with other secreted glycoproteins, N‐linked glycosylation of Bβ and γ chains commences in the ER and is completed in Golgi organelles. Sulfation and phosphorylation occur at post‐ER stages of the secretory process. Since some ER chaperones coisolate with nascent fibrinogen chains they have been implicated in assisting chain assembly. Studies with recombinant systems, using deletion and substitution mutants, indicate that initial chain assembly depends on hydrophobic interactions present in the C‐terminal half of the coil‐coil domains and that inter‐ and intra‐disulfide bonds that stabilize fibrinogen are needed to complete chain assembly. Not all the chains that are synthesized are assembled into fibrinogen and the unassembled chains are not secreted. HepG2 cells contain surplus Aα and γ chains that accumulate as free γ chains and as an Aα‐γ complex. Aα‐γ is degraded by lysosomes whereas the γ chain is degraded by the proteasome‐ubiquitin system. Studies with expression of single chains by COS cells confirm that γ and Bβ are hydrolyzed by proteasomes and indicate that Aα is degraded partially both by lysosomes and proteasomes. The role of surplus chains in regulating fibrinogen assembly is not understood but overexpression of any one chain, elicited by transfection of HepG2 cells, results in the upregulation of the other two genes, increased fibrinogen synthesis and secretion, and maintenance of surplus intracellular Aα and γ chains. HepG2 cells, programmed in this manner to increase basal fibrinogen expression, have higher HMG‐CoA reductase mRNA levels, enhanced cholesterol and cholesterol ester synthesis, and increased secretion of apolipoprotein B (apoB). Overexpression of basal levels of fibrinogen does not affect synthesis of other acute phase proteins. Enhanced secretion of apoB is due to diminished degradation of nascent apoB by proteasomes and not to increased expression. Increased secretion of apoB is associated with increased basal expression of fibrinogen and is not affected when fibrinogen expression is stimulated by interleukin‐6. In HepG2 cells, a feedback mechanism exists and extracellular sterols specifically downregulate expression of the three fibrinogen genes. These studies link, at the cellular level, basal fibrinogen expression with lipid metabolism.