Table 1 List of serine proteinase inhibitors which either occur as part of other functional proteins or show other functions*

Inhibitor

Serine proteinase inhibition

Other function

STI (Kunitz)

Nodulin1

(found in winged bean nodules when senescence begins)

Inhibits trypsin and other endogenous proteinases.

Interacts with bacteroids and is believed to be involved in nodule senescence.

 

Miraculin2

(found in red berries of Richadella dulcifica)

Inhibitory function not known, ~40% sequence identity to STI.

Changes sour taste into sweet taste.

Wheat amylase / Subtilisin inhibitor (WASI)3

Inhibits subtilisin

Inhibits a -amylase. Endogenous

Rice amylase / Subtilisin inhibitor (RASI)4

Inhibits subtilisin

Inhibits a -amylase. Endogenous

Barley amylase / Subtilisin inhibitor (BASI)5

Inhibits subtilisin

Inhibits a -amylase.

Endogenous

Cathepsin D inhibitor

(PDI)6 (found in potato)

Inhibits trypsin

Inhibits cathepsin D, an aspartate proteinase.

Novel inhibitor of cathepsin D (NID)7

Inhibits trypsin

Inhibits cathepsin D

Wound induced Asp proteinase (CDI)8

Inhibits trypsin

Inhibits cathepsin D

Potato cysteine proteinase inhibitor (PCPI)9

Inhibitory function not known, 23% sequence identity to STI

Inhibits cathepsin L, a cysteine proteinase

Winged bean albumin (WBA-1)10

 

Inhibitory activity not known

Presumably a storage protein

BPTI (Kunitz)

BPTI11-13

 

Strong inhibitor of trypsin and kallikrein. Also inhibits many other serine proteinases

 

BPTI and its fragments 1-15, 18-39, and 40-58 have been shown to posses antibacterial activity against many gram positive and gram negative bacteria.

Inhibits adenovirus cysteine proteinase. Binding constant with recombinant virus proteinase is 5 x 105. The inhibitory site is believed to be different than that for trypsin.

Inhibits Ca++ activated K+ channel. The a subunit of KCa channel contains a conserved proteinase inhibitor binding site.

APPI14, 15

Several isoforms of Amyloid Precursor Protein (APP). Two isoforms, APP (751) and APP (&) contain a Kunitz type inhibitor domain, APPI.

 

APPI is a strong inhibitor of trypsin. It also inhibits many other serine proteinases.

 

 

 

 

 

APPs function as neuronal receptors believed to be involved in intracellular signaling pathway through GTP binding protein, Go. In Alzheimers disease, a fragment of APP, amyloid b protein is deposited in various brain tissues.

Calcicludine16

(found in the venom of green mamba snake)

Inhibitory activity not known.

Specifically blocks most of the high threshold Ca++ channels.

Dendrotoxins 17, 18

(found in the venom of several mamba snakes) examples: " -dendrotoxin, toxin I, toxin K, and toxin E.

Inhibit trypsin and kallikrein.

Block some neuronal K+ channels and augment release of neurotransmitters.

B chain of b 1 and b 2 bungarotoxins19 (found in the venom of Bungrus multicinctus)

Inhibitory activity not known.

Bungarotoxin have two chains: A & B. They block neuromuscular transmission. A chain is a phospholipase.

Bikunin20, 21

Inhibits trypsin, chymotrypsin, PPE, and HLE.

Bikunin is co-translated with a 1 microglobulin which belongs to lipocalin superfamily. This resulting precursor undergoes post-translational processing to produce a 1 microglobulin and bikunin. Bikunin then becomes covalently associated with one or two chains related to multioxidase famaily to form either Pre " I or Ia I.

Type VII and Type VI22, 23 (a 3 chain) collagen contain a Kunitz like inhibitor domain at their C-terminus.

No inhibitory activity reported. However, a single mutation at P1’ (DA) makes it a weak inhibitor of trypsin. A triple mutant (P3TP; P1’DA; P2’FR) is a very strong inhibitor of trypsin.

Type VII is a component of anchoring fibrils while type VI acts as a cell binding protein.

BPTI/Bikunin/Trypstatin24

See above

Inhibit binding of HIV glycoprotein gp 120 with tryptase TL.

 

Kazal

Ovoinhibitor13

 

 

 

 

 

Inhibits a variety of serine proteinases. Has seven inhibitory domains.

 

Inhibits Ca++ activated K+ channel. The a subunit of KCa channel contains a conserved proteinase inhibitor binding site.

Follistatin25

Inhibitory activity not known.

Modulates the action of several members of the family of transforming growth factor b .

Agrin26

Among others, contains nine follistatin-like domains

Some of the Kazal/follistatin domains in agrin inhibit CARL and HLE (Fisher, J. – personal communication)

Associated with basal membranes of several tissues such as synaptic basal lamina of neuromuscular junction.

BM-4027

(SPARC or Osteonectin) Among others, contains a follistatin-like domain

Inhibitory activity not known.

Antiadhesive secreted glycoprotein. Believed to be involved in tissue remodelling.

PEC-6028

Inhibits trypsin (unpublished results)

Structurally similar to cholecystokinin release factor. It is also known to suppress glucose induced insulin secretion.

Thrombospondin 129

(a multidomain glycoprotein from platelets)

Contains two Kazal type sequences. Inhibits HLE and cathepsin G.

Believed to be involved in tissue development and remodeling.

PSTI II30


Inhibits trypsin.

Shows cholecystokinin releasing activity. A peptide (monitor peptide) similar to PSTI and showing strong cholecystokinin release activity has also been isolated.

Rhodniin31

First domain of rhodniin strongly inhibits thrombin.

Second domain of rhodniin is believed to be non-inhibitory but binds to the fibrinogen recognition site on thrombin.

 

SSI

SSI32 and other similar inhibitors such as API-2c & PS

 

Strong inhibitors of subtilisins and many other serine proteinases.

 

 

Inhibit Streptomyces griseus metallo endopeptidase. The serine proteinase and the metallo endopeptidase inhibitory sites overlap.

Cereal family

RBI33

 

Inhibits trypsin

 

Inhibits " -amylase. Many other members of this family inhibit " -amylase but they don’t have known proteinase inhibitory activity.

 

Grasshopper family

Pacifastin34

(found in the plasma of freshwater Crayfish, Pacifastacus Leniusculus

 

Inhibits Chymotrypsin, trypsin, PPE and a proteinase from Crayfish haemolymph.

 

Composed of a light chain with 9 inhibitory domains and a heavy chain. The heavy chain is structurally and functionally related to transferrin and has at least one presumably two iron binding sites.

 

Chelonianin

SLPI35

 

Inhibits many serine proteinases

 

Inhibits HIV-1 infectivity of monocytes and T lymphocytes.

*Only selected examples are presented here. Many inhibitors are known to serve as storage proteins. Some examples are ovomucoid and many plant inhibitor families such as Kunitz (STI), cereal, BBI etc.

References:

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2. Theerasilp, S., Hitotsuya, H., Nakajo, S., Nakaya, K., Nakamura, Y., and Kurihara, Y. (1989) Complete amino acid sequence and structure characterization of the taste-modifying protein, miraculin. J. Biol. Chem., 264, 6655-6659.

3. Mundy, J., Hejgaard, J., and Svendsen I. (1984) Characterization of a bifunctional wheat inhibitor of endogenous a -amylase and subtilisin. FEBS Letters, 167, 210-.

4. Ohtsubo, K.-I. and Richardson, M. (1992) The amino acid sequence of a 20 kDa bifunctional subtilisin / a-amylase inhibitor from bran of rice (Oryza sativa L.) seeds. FEBS Letters, 309, 68-72.

5. Vallee, F., Kadziola, A., Bourne, Y., Juy, M., Rodenburg, K.W., Svensson, B., and Haser, R. (1998) Barley a-amylase bound to its endogenous protein inhibitor BASI: Crystal structure of the complex at 1.9 D resolution. Structure, 6, 649-659.

6. Mares, M., Meloun, B., Pavlik, M., Kostka, V., and Baudys, M. (1989) Primary structure of cathepsin D inhibitor from potatoes and its structure relationship to soybean trypsin inhibitor family. FEBS Letters, 251, 94-98.

7. Ritonja, A., Krizaj, I., Mesko, P, Kopitar, M,. Lucovnik, P., Otrukelj, B., Pungercar, J., Buttle, D.J., Barrett, A.J., and Turk, V. (1990) The amino acid sequence of a novel inhibitor of cathepsin D from potato. FEBS Letters, 267, 13-15.

8. Hildmann, T., Ebneth, M., Pena-Cortes, H., Sanchez-Serrano, J. J., Willmitzer, L., and Prat, S. (1992) General roles of abscisic and jasmonic acids in gene activation as a result of mechanical wounding. The Plant Cell, 4, 1157-1170.

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16. Schweitz, H., Heurteaux, C., Bois, P., Moinier, D., Romey, G., and Lazdunski, M. (1994) Calcicludine, a venom peptide of the Kunitz type protease inhibitor family, is a potent blocker of high-threshold Ca2+ channels with a high affinity for L-type channels in cerebellar granule neurons. Proc. Nat. Acad. Sci., 91, 878-882.

17. Skarzynski, T. (1992) Crystal structure of a-dendrotoxin from the green mamba venom and its comparison with the structure of bovine pancreatic trypsin inhibitor. J. Mol. Biol., 224, 671-683.

18. Marshall, D.L. and Haravey, A.L. (1992) Protease inhibitor homologues of dendrotoxin do not bind to dendrotoxin acceptors on synaptosomal membranes or facilitate neuromuscular transmission. Biol. Chem. Hoppe-Seyler, 373, 707-714.

19. Kondo, K., Toda, H., and Narita, K. (1982) Amino acid sequence of b-bungarotoxin from Bungarus multicinctus venom. The amino acid substitutions in the B chains. J. Biochem., 91, 1519-1530.

20. Lindqvist, A., and Akerström, B. (1996) Bovine a1-microglobulin / bikunin. Isolation and characterization of liver cDNA and urinary a1-microglobulin. Biochim. Biophys. Acta, 1306, 98-106.

21. Itoh, H., Ide, H., Ishikawa, N., and Nawa, Y. (1994) Mast cell protease inhibitor, trypstatin, is a fragment of inter-a-trypsin inhibitor light chain. J. Biol. Chem., 269, 3818-3822.

22. Kohfeldt, E., Göhring, W., Mayer, U. Zweckstetter, M., Holak, T.A., Chu, M.-L., and Timpl, R. (1996) Conversion of the Kunitz type module of collagen VI into a highly active trypsin inhibitor by site-directed mutagenesis. Eur. J. Biochem., 238, 333-340.

23. Greenspan, D. S. (1993) The carboxy terminal half of type VII collagen including the NC-2 domain and intron/exon organization of the corresponding region of the COL7A1 gene. Hum. Mol. Genet. 2, 273-278.

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27. Hohenester, E., Maurer, P., and Timpl, R. (1997) Crystal structure of a pair of follistatin-like and EF-hand calcium binding domains in BM-40. EMBO J., 16, 3778-3786.

28. Agerberth, B., Söderling-Barros, J., Jörnvall, H., Chen, S.-W., Östensen, C.-G., Efendic, S., and Mutt, V. (1989) Isolation and characterization of a 60 residue intestinal peptide structurally related to the pancreatic secretory type of trypsin inhibitor: Influence on insulin secretion. Proc. Nat. Acad. Sci., 86, 8590-8594.

29. Hogg, P.J., Jimenez, B. M., and Chesterman, C. N. (1994) Identification of possible inhibitory reactive centers in thrombospondin 1 that may bind cathepsin G and neutrophil elastase. Biochemistry, 33, 6531-6537.

30. Tsuzuki, S., Fushiki, T., Kondo, A., Murayama, H., and Sugimoto, E. (1991) Effect of a high-protein diet on the gene expression of a trypsin-sensitive, cholecystokinin-releasing peptide (monitor peptide) in the pancreas. Eur. J. Biochem, 199, 245-252.

31. van de Locht, A., Lamba, D., Bauer, M., Huber, R., Friedrich, T., Kröger, B., Höffken, W., and Bode, W. (1995) Two heads are better than one: Crystal structure of the insect derived double domain Kazal inhibitor rhodniin in complex with thrombin. EMBO J., 14, 5149-5157.

32. Kumazaki, T., Kajiwara, K., Kojima, S., Miura, K., and Ishii, S. (1993) Interaction of Streptomyces subtilisin inhibitor (SSI) with Streptomyces griseus metallo-endopeptidase II (SGMP II). J. Biochem., 114, 570-575.

33. Strobl, S., Muhlhahn, P., Bernstein, R., Wiltscheck, R., Maskos, K., Wunderlich, M., Huber, R., Glockshuber, R., and Holak, T.A. (1995) Determination of the three- dimensional structure of the bifunctional a-amylase / trypsin inhibitor from ragi seeds by NMR spectroscopy. Biochemistry, 34, 8281-8293.

34. Liang, Z., Sottrup-Jensen, L., Aspán, A., Hall, M., and Söderhall, K. (1997) Pacifastin, a novel 155-kDa heterodimeric proteinase inhibitor containing a unique transferrin chain. Proc. Nat. Acad. Sci., 94, 6682-6687.

35. McNeely, T. B., Dealy, M., Dribbs, D.J., Orenstein, J. M., Eisenberg, S.P., and Wahl, S. M. (1995) Secretory leukocyte protease inhibitor: A human saliva protein exhibiting anti-human immunodeficiency virus activity in vitro. J. Clin. Invest., 96, 456-464.