Tissue prothrombin. Universal distribution in smooth muscle.
Immunohistochemical analysis of surgically obtained porcine tissue samples reveals ubiquitous staining for prothrombin in organs rich in smooth muscle content and universal staining of smooth muscle in tissue vasculature. The native character of tissue prothrombin is verified first by chromogenic substrate hydrolysis and hirudin inhibition after incubation of tissue extracts with taipan snake venom and phospholipid. Western analysis of tissue extracts confirms the native zymogen molecular weight. In addition, prothrombin purified in good yield from porcine uterus is activated by Echis carinatus venom which, like taipan venom, is 4-carboxyglutamic acid-sensitive. After correction for blood (gross heme) and interstitial fluid (albumin), excess functional prothrombin is observed in extracts of tissues having abundant smooth muscle. In contrast with factor X, the yield of prothrombin purified from porcine uterus greatly exceeds that attributable to contamination by whole blood. Northern blot analysis from selected bovine tissues extracted for polyadenylated messenger RNA is equivocal for prothrombin mRNA with the exception of liver, which is positive. It is concluded that functionally intact prothrombin is widely distributed among tissues owing to smooth muscle content, although the mechanism of emplacement and physiologic significance of prothrombin in these tissues remains unclear.
McBane
R D
RD
Section of Hematology Research, Mayo Clinic and Foundation for Education and Research, Rochester, Minn. 55905, USA.
Miller
R S
RS
Hassinger
N L
NL
Chesebro
J H
JH
Nemerson
Y
Y
Owen
W G
WG
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D004789:Enzyme Activation / Q000187:drug effects; D005110:Extracellular Space / Q000737:chemistry; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009130:Muscle, Smooth / Q000737:chemistry*; D009928:Organ Specificity; D010446:Peptide Fragments / Q000032:analysis; D011516:Prothrombin / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis; D012910:Snake Venoms / Q000494:pharmacology; D013552:Swine; D014599:Uterus / Q000737:chemistry
D010446:Peptide Fragments; D012333:RNA, Messenger; D012910:Snake Venoms; C045879:prothrombin fragment 1.2; D011516:Prothrombin