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Title: Recombinant human fibrinogen and sulfation of the. gamma. prime chain

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00103a004· OSTI ID:5603784
; ; ;  [1];  [2]
  1. Univ. of Washington, Seattle (United States)
  2. ZymoGenetics, Inc., Seattle, WA (United States)

Human fibrinogen and the homodimeric {gamma}{prime}-chain-containing variant have been expressed in BHK cells using cDNAs coding for the {alpha},{beta}, and {gamma} (or {gamma}{prime}) chains. The fibrinogens were secreted at levels greater than 4 {mu}g (mg of total cell protein){sup {minus}1}day{sup {minus}1} and were biologically active in clotting assays. Recombinant fibrinogen containing the {gamma}' chain incorporated {sup 35}SO{sub 4} into its chains during biosynthesis, while no incorporation occurred in the protein containing the {gamma} chain. The identity of the sulfated {gamma}{prime} chain was verified by its ability to form dimers during clotting. In addition, carboxypeptidase {Upsilon} digestion of the recombinant fibrinogen containing the {gamma}{prime} chain released 96% of the {sup 35}S label from the sulfated chain, and the radioactive material was identified as tyrosine O-sulfate. These results clarify previous findings of the sulfation of tyrosine in human fibrinogen.

OSTI ID:
5603784
Journal Information:
Biochemistry; (United States), Vol. 30:39; ISSN 0006-2960
Country of Publication:
United States
Language:
English