Primary structures of guinea pig high- and low-molecular-weight kininogens

  • Semba U
  • Umeda Y
  • Shibuya Y
 et al. 
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Abstract

Guinea pig high-molecular-weight and low-molecular-weight (HMW and LMW) kininogen cDNA were amplified from liver mRNA by RT-PCR. Their nucleotide sequences were analyzed and deduced to amino acid sequences. The HMW kininogen, composed of 607 amino acid residues with a 18-residue signal sequence, possessed the cysteine protease inhibitor domains I and II, the bradykinin domain, the histidine-rich region, and the prekallikrein-binding region. The amino acid sequence preceding the bradykinin domain was found not to be -Leu-Met-Lys- but -Leu-Thr-Arg-. Therefore, kallidin (Lys-bradykinin) and Met-kallidin are not liberated from the guinea pig kininogens. We purified the HMW kininogen protein from plasma and prepared the kinin-free form using guinea pig plasma kallikrein. Although the amino-terminal of the HMW kininogen was modified, the 25 amino-terminal residues of the light chain of the kinin-free kininogen corresponded to the deduced sequence just after the bradykinin moiety of the HMW kininogen. With regard to the LMW kininogen, the nucleotide sequence down to T1200as well as the amino acid sequence till Thr382was identical to that of the HMW kininogen. We also examined the localization of the guinea pig kininogen gene on the prometaphase lymphocyte chromosomes by fluorescence in situ hybridization method. Two pair signals were observed on a pair of homologous chromosomes, each of which is composed of two chromatids. Based on these findings, we concluded that HMW and LMW kininogens are produced from the single kininogen gene in guinea pig as in the cases of the other mammalian species reported so far. © 2004 Elsevier B.V. All rights reserved.

Author-supplied keywords

  • 4′,6-diamidino-2- phenylindole
  • Amino acid sequence
  • Bradykinin
  • DAPI
  • DIG
  • FISH
  • Guinea pig
  • HMW
  • High-molecular-weight kininogen
  • Kininogen gene
  • LMW
  • Low-molecular-weight kininogen
  • digoxigenin
  • fluorescence in situ hybridization
  • high-molecular-weight

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Authors

  • Umeko Semba

  • Yasuko Umeda

  • Yoko Shibuya

  • Hiroaki Okabe

  • Sumio Tanase

  • Tetsuro Yamamoto

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