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. 1997 Jul 15;325(Pt 2):527–531. doi: 10.1042/bj3250527

Purification and characterization of three distinct types of phospholipase A2 inhibitors from the blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus.

N Ohkura 1, H Okuhara 1, S Inoue 1, K Ikeda 1, K Hayashi 1
PMCID: PMC1218591  PMID: 9230137

Abstract

Three distinct types of phospholipase A2 (PLA2) inhibitory proteins (PLIalpha, PLIbeta, and PLIgamma) were isolated from the blood plasma of the Chinese mamushi, Agkistrodon blomhoffii siniticus. PLIalpha is an inhibitor that we have already purified and whose amino acid sequence we have already determined [Ohkura, Inoue, Ikeda and Hayashi (1993) J. Biochem. (Tokyo) 113, 413-419]. It inhibited selectively the group-II acidic PLA2s from Crotalidae venom. PLIbeta was a 160-kDa glycoprotein having a trimeric structure composed of 50-kDa subunits. The amino acid sequence of the first 30 amino acids of the N-terminal part of the 50-kDa subunit was determined and found to have no significant homology to that of known proteins. PLIbeta was a selective inhibitor against the group-II basic PLA2s from Crotalidae venom. Some amino acid residues located in or close to the interfacial binding surface of the group-II basic PLA2s were suggested to be involved in selective binding to PLIbeta. PLIgamma was a 100-kDa glycoprotein containing 25-kDa and 20-kDa subunits and inhibited all of the PLA2s investigated equally, including Elapidae venom PLA2s (group I), Crotalidae and Viperidae venom PLA2s (group II) and honey-bee PLA2 (group III). From the N-terminal sequences of the two subunits, PLIgamma was found to be the same type of PLI that had been purified from Thailand cobra plasma.

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Selected References

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  1. Cavins J. F., Friedman M. An internal standard for amino acid analyses: S-beta-(4-pyridylethyl)-L-cysteine. Anal Biochem. 1970 Jun;35(2):489–493. doi: 10.1016/0003-2697(70)90211-3. [DOI] [PubMed] [Google Scholar]
  2. Chen Y. C., Maraganore J. M., Reardon I., Heinrikson R. L. Characterization of the structure and function of three phospholipases A2 from the venom of Agkistrodon halys pallas. Toxicon. 1987;25(4):401–409. doi: 10.1016/0041-0101(87)90073-0. [DOI] [PubMed] [Google Scholar]
  3. Forst S., Weiss J., Blackburn P., Frangione B., Goni F., Elsbach P. Amino acid sequence of a basic Agkistrodon halys blomhoffii phospholipase A2. Possible role of NH2-terminal lysines in action on phospholipids of Escherichia coli. Biochemistry. 1986 Jul 29;25(15):4309–4314. doi: 10.1021/bi00363a020. [DOI] [PubMed] [Google Scholar]
  4. Fortes-Dias C. L., Lin Y., Ewell J., Diniz C. R., Liu T. Y. A phospholipase A2 inhibitor from the plasma of the South American rattlesnake (Crotalus durissus terrificus). Protein structure, genomic structure, and mechanism of action. J Biol Chem. 1994 Jun 3;269(22):15646–15651. [PubMed] [Google Scholar]
  5. Heinrikson R. L., Krueger E. T., Keim P. S. Amino acid sequence of phospholipase A2-alpha from the venom of Crotalus adamanteus. A new classification of phospholipases A2 based upon structural determinants. J Biol Chem. 1977 Jul 25;252(14):4913–4921. [PubMed] [Google Scholar]
  6. Inoue S., Kogaki H., Ikeda K., Samejima Y., Omori-Satoh T. Amino acid sequences of the two subunits of a phospholipase A2 inhibitor from the blood plasma of Trimeresurus flavoviridis. Sequence homologies with pulmonary surfactant apoprotein and animal lectins. J Biol Chem. 1991 Jan 15;266(2):1001–1007. [PubMed] [Google Scholar]
  7. Inoue S., Shimada A., Ohkura N., Ikeda K., Samejima Y., Omori-Satoh T., Hayashi K. Specificity of two types of phospholipase A2 inhibitors from the plasma of venomous snakes. Biochem Mol Biol Int. 1997 Mar;41(3):529–537. doi: 10.1080/15216549700201551. [DOI] [PubMed] [Google Scholar]
  8. Joubert F. J., Taljaard N. Purification, some properties and amino-acid sequences of two phospholipases A (CM-II and CM-III) from Naja naja kaouthia venom. Eur J Biochem. 1980 Dec;112(3):493–499. doi: 10.1111/j.1432-1033.1980.tb06112.x. [DOI] [PubMed] [Google Scholar]
  9. Kini R. M., Kawabata S. I., Iwanaga S. Comparison of amino terminal region of three isoenzymes of phospholipases A2 (TFV PL-Ia, TFV PL-Ib, TFV PL-X) from Trimeresurus flavoviridis (habu snake) venom and the complete amino acid sequence of the basic phospholipase, TFV PL-X. Toxicon. 1986;24(11-12):1117–1129. doi: 10.1016/0041-0101(86)90138-8. [DOI] [PubMed] [Google Scholar]
  10. Kogaki H., Inoue S., Ikeda K., Samejima Y., Omori-Satoh T., Hamaguchi K. Isolation and fundamental properties of a phospholipase A2 inhibitor from the blood plasma of Trimeresurus flavoviridis. J Biochem. 1989 Dec;106(6):966–971. doi: 10.1093/oxfordjournals.jbchem.a122983. [DOI] [PubMed] [Google Scholar]
  11. Kondo K., Zhang J., Xu K., Kagamiyama H. Amino acid sequence of a presynaptic neurotoxin, agkistrodotoxin, from the venom of Agkistrodon halys Pallas. J Biochem. 1989 Feb;105(2):196–203. doi: 10.1093/oxfordjournals.jbchem.a122639. [DOI] [PubMed] [Google Scholar]
  12. Oda N., Ogawa T., Ohno M., Sasaki H., Sakaki Y., Kihara H. Cloning and sequence analysis of cDNA for Trimeresurus flavoviridis phospholipase A2, and consequent revision of the amino acid sequence. J Biochem. 1990 Nov;108(5):816–821. doi: 10.1093/oxfordjournals.jbchem.a123286. [DOI] [PubMed] [Google Scholar]
  13. Ohkura N., Inoue S., Ikeda K., Hayashi K. Isolation and amino acid sequence of a phospholipase A2 inhibitor from the blood plasma of Agkistrodon blomhoffii siniticus. J Biochem. 1993 Apr;113(4):413–419. doi: 10.1093/oxfordjournals.jbchem.a124060. [DOI] [PubMed] [Google Scholar]
  14. Ohkura N., Inoue S., Ikeda K., Hayashi K. Isolation and characterization of a phospholipase A2 inhibitor from the blood plasma of the Thailand cobra Naja naja kaouthia. Biochem Biophys Res Commun. 1994 Apr 29;200(2):784–788. doi: 10.1006/bbrc.1994.1519. [DOI] [PubMed] [Google Scholar]
  15. Ohkura N., Inoue S., Ikeda K., Hayashi K. The two subunits of a phospholipase A2 inhibitor from the plasma of Thailand cobra having structural similarity to urokinase-type plasminogen activator receptor and LY-6 related proteins. Biochem Biophys Res Commun. 1994 Nov 15;204(3):1212–1218. doi: 10.1006/bbrc.1994.2592. [DOI] [PubMed] [Google Scholar]
  16. Perales J., Villela C., Domont G. B., Choumet V., Saliou B., Moussatché H., Bon C., Faure G. Molecular structure and mechanism of action of the crotoxin inhibitor from Crotalus durissus terrificus serum. Eur J Biochem. 1995 Jan 15;227(1-2):19–26. doi: 10.1111/j.1432-1033.1995.tb20355.x. [DOI] [PubMed] [Google Scholar]
  17. Radvanyi F., Jordan L., Russo-Marie F., Bon C. A sensitive and continuous fluorometric assay for phospholipase A2 using pyrene-labeled phospholipids in the presence of serum albumin. Anal Biochem. 1989 Feb 15;177(1):103–109. doi: 10.1016/0003-2697(89)90022-5. [DOI] [PubMed] [Google Scholar]
  18. Renetseder R., Brunie S., Dijkstra B. W., Drenth J., Sigler P. B. A comparison of the crystal structures of phospholipase A2 from bovine pancreas and Crotalus atrox venom. J Biol Chem. 1985 Sep 25;260(21):11627–11634. [PubMed] [Google Scholar]
  19. Takasaki C., Yutani F., Kajiyashiki T. Amino acid sequences of eight phospholipases A2 from the venom of Australian king brown snake, Pseudechis australis. Toxicon. 1990;28(3):329–339. doi: 10.1016/0041-0101(90)90068-i. [DOI] [PubMed] [Google Scholar]
  20. Tomoo K., Ohishi H., Ishida T., Inoue M., Ikeda K., Aoki Y., Samejima Y. Revised amino acid sequence, crystallization, and preliminary x-ray diffraction analysis of acidic phospholipase A2 from the venom of Agkistrodon halys blomhoffii. J Biol Chem. 1989 Feb 25;264(6):3636–3638. [PubMed] [Google Scholar]
  21. Tsai I. H., Wu S. H., Lo T. B. Complete amino acid sequence of a phospholipase A2 from the venom of Naja naja atra (Taiwan cobra). Toxicon. 1981;19(1):141–152. doi: 10.1016/0041-0101(81)90126-4. [DOI] [PubMed] [Google Scholar]

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