Characterization of guinea-pig high-molecular-weight kininogen as multi-functional molecule.

Abstract

High-molecular-weight (HMW) kininogen was purified from guinea-pig plasma by measuring its ability to correct the prolonged clotting time in human HMW kininogen deficient plasma (Fitzgerald trait). The purified HMW kininogen demonstrated a homogeneous band in disc gel electrophoresis in the presence of sodium dodecyl sulfate under reducing or non-reducing conditions with an apparent molecular weight of 100,000. Kinin released from HMW kininogen by treatment with guinea-pig plasma kallikrein was identified as bradykinin by reverse-phase HPLC and amino-acid analysis. The capacity of HMW kininogen as a thiol-proteinase inhibitor was realized by its dose-dependent inhibitory activity to papain. The Ki value for papain was estimated to be 42 pM. The kinin-free HMW kininogen maintained the inhibitor and clotting-factor activities with similar capacities to those of the HMW kininogen molecule. Heavy chain (H-chain) and light chain (L-chain) of HMW kininogen were prepared from reduced and alkylated kinin-free HMW kininogen by HPLC. The S-alkylated H-chain, but not L-chain, demonstrated the inhibitor activity with the Ki value 6.9 nM for papain, whereas the S-alkylated L-chain, but not H-chain, maintained the clotting activity one-third of the capacity of HMW kininogen. Specific antibodies recognized HMW kininogen, but also a probable low-molecular-weight kininogen(s) with an apparent molecular weight of 60,000 in the guinea-pig plasma. All of these properties are consistent with the reports on human, bovine and rat HMW kininogen.

• Authors: T Yamamoto
• Journal: Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 914 Issue 3 Pg. 259-74 (Aug 21 1987) ISSN: 0006-3002 [Print] Netherlands
• PMID: 3113483 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Amino Acids
• Kininogens
• Kinins
• Macromolecular Substances
• Papain

Topics

• Amino Acids (analysis)
• Animals
• Blood Coagulation
• Enzyme-Linked Immunosorbent Assay
• Female
• Guinea Pigs
• Immunodiffusion
• Immunoelectrophoresis
• Kinetics
• Kininogens (blood, isolation & purification, physiology)
• Kinins (analysis)
• Macromolecular Substances
• Male
• Molecular Weight
• Papain (antagonists & inhibitors)