EMAIL THIS PAGE TO A FRIEND

Biochemistry

Characterization of bovine prothrombin mRNA and its translation product.


PMID 6326805

Abstract

Prothrombin mRNA has been enriched 20-60-fold by using specific immunoadsorption of bovine liver polysomes. The enriched mRNA was translated in a cell-free protein synthesizing system derived from rabbit reticulocytes, and the radiolabeled translation product was isolated by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The radiolabeled prothrombin synthesized in the cell-free system was then subjected to automated Edman degradation and shown to contain a leader sequence of at least 30 residues that was rich in leucine, phenylalanine, and alanine. In order to fully characterize the leader sequence for prothrombin, a bovine liver cDNA library was constructed containing DNA inserts of over 1000 base pairs. Two cDNA clones coding for bovine prothrombin were isolated from the library and their nucleotide sequences determined. A leader sequence of 43 amino acids was predicted from the sequence of the cDNA, and the first 30 residues were in agreement with the partial sequence obtained by the cell-free protein synthesizing system. From the amino acid sequence of the leader sequence, it is proposed that bovine prothrombin is synthesized with a prepro leader sequence starting with a methionine residue at position -43. The amino acid sequence of the mature prothrombin molecule circulating in plasma was also predicted from the cDNA and shown to be in good agreement with that determined previously by conventional amino acid sequence analysis.

Related Materials

Product #

Image

Description

Molecular Formula

Add to Cart

T4648 Thrombin from bovine plasma, lyophilized powder, 40-300 NIH units/mg protein (biuret)
T7513 Thrombin from bovine plasma, lyophilized powder, ≥2,000 NIH units/mg protein (E1%/280 = 19.5)
T6634 Thrombin from bovine plasma, lyophilized powder, 600-2,000 NIH units/mg protein (biuret)
T9549 Thrombin from bovine plasma, powder, cell culture tested, ≥1,500 NIH units/mg protein (E1%/280 = 19.5)
T9000 Thrombin from bovine plasma, buffered aqueous solution, ≥125 NIH units/mg protein (biuret)
T7201 Thrombin from bovine plasma, lyophilized powder, ≥2,000 NIH units/mg protein (E1%/280 = 19.5)
T7326
Thrombin from bovine plasma, ≥60 NIH units/mg protein (biuret)
T6884
Thrombin from human plasma, lyophilized powder, ≥2,000 NIH units/mg protein (E1%/280, 18.3)
T7009
Thrombin from human plasma, lyophilized powder, ≥1,000 NIH units/mg protein (E1%/280, 18.3)
T1063
Thrombin from human plasma, lyophilized powder, ≥2800 NIH units/mg protein (E1%/280, 18.3)
T8885
Thrombin from human plasma, lyophilized powder, Suitable for routine use in the thrombin time test
T4393
Thrombin from human plasma, lyophilized powder, 1500-3500 NIH units/mg protein (E1%/280, 18.3), cell culture tested
T7572
Thrombin from human plasma, 400-1000 NIH units/mg protein
T5772 Thrombin from rat plasma, lyophilized powder, ≥1,000 units/mg protein (biuret)
T9326 Thrombin human, BioUltra, recombinant, expressed in HEK 293 cells, aqueous solution, ≥95% (SDS-PAGE)