Biochemistry of Proteins

PROTEASE INHIBITORS

See also: Detergents - Dialysis - Protein dosage (detergent compatibility)

 

Uptima offers several protease inhibitors for requirements of :

  • Protein solubilization, extraction and purification

  • Protein storage, protein analysis

  • Biochemistry of proteins

  1. Proteolysis, general information

  2. Protease inhibitors selection guide

  3. Protease inhibitors from Uptima

    Ac-Asp-Glu-Val-(Haldehyde)

    Ac-Tyr-Val-Ala-Asp-H(aldehyde)

    Ac-Tyr-Val-Lys-Asp-H(aldehyde)

    CA-074

    CA-074 Me

    Chymostatin

    E-64

    E-64-c

    E-64-d

    AEBSF

    Antipain HCl

    Aprotinin

    Bestatin

    Biotinyl-Asp-Glu-Val-Asp-H(ald.)

    Elafin (Human)

    Leupeptin

    Pepstatin

    Phosphoramidon

    PMSF

    Trypsin inhibitor

    EDTA

    EGTA

    Z-Asp-CH2-DCB

    Z-Ileu-Glu(OBut)-Ala-Leu-H

    Z-Leu-Leu-H

    Z-Leu-Leu-Leu-H

    Z-Leu-Leu-Nva-H

Proteolysis, general information

Proteases are ubiquitous enzymes in every cells of all organisms. They are released during disruption of cells, and quickly degrade any protein. When one wants to recover or detect a given protein, protease inhibition is thus required to save the yield and the quality of the protein. Protease inhibitors are also useful to protect proteins during further analysis (western blotting, immunoprecipitation, bioassays, reporter gene detection…), purification steps (chromatography), and storage.

Uptima recommends that protease inhibitors with broad specific activity are added to all stock buffers and solutions, and not only in initial purification step, because most inhibitors have reversible activity. A classic complete association is 3mg antipain, 0.5mg Bestatin, 1mg chymostatin, 3mg E-64, 0.5mg pepstatin, 3mg phosphoramidon, 20mg PefablocSC, 0.5mg aprotinine and 10mg EDTA. EDTA may be avoided if metalloproteases are not a concern, notably in applications where divalent metals are needed (polyHist tagged recombinant protein affinity purification, enzymes with cofactors…)

Protease inhibitors selection guide

Different types of proteases exist. Serine proteases are present in almost all cells, dominating in plants with cysteine proteases or in bacteria with metalloproteases. Occasionally, aspartic proteases can interfere too in animal tissues isolations notably when a low pH is required.

The table below indicates most important general inhibitors dedicated to these protease types, and inhibiting notably non-specific proteases that are commonly found . The table gives also interesting inhibitors for specific applications, in example for specific proteases that are involved in physiological pathways of cells and living organisms.

For best results, choose Uptima high quality protease inhibitors

Proteases

General Inhibitors

Specific Inhibitors / uses and other inhibitors

Serine proteases

-

-

Chymotrypsin

Æ AEBSF is a superior serine protease inhibitor, with far lower toxicity than conventional PMSF and DFP

 

See also :

Aprotinin, E-64, Leupeptin, PMSF

Chymostatin, Soybean Trypsin Inhibitor

Kallikrein

-

Plasmin

-

Subtilisin

PMSF

Thrombin

-

Trypsin

Soybean Trypsin Inhibitor

Cysteine proteases

-

-

Calpain

 

Æ Antipain and E64

See also : Leupeptin

UP831930

Cathepsin B, H, L, S

CA-074

Papain

Chymostatin, PMSF

Metalloproteases

-

-

Ca+ metalloproteases

 

Æ EDTA

EGTA

MetalloEndoProteinases

Phosphoramidon

Aspartic acid proteases

-

-

Cathepsin D

 

Æ Pepstatin

---

Pepsin

Renin

Others, specific proteases

-

-

Aminopeptidase

 

 

 

Æ Bestatin

Bestatin

Apopain

UP827070, UP827100

Collagenase

Phosphoramidon

Elastase

Elastase, Soybean Trypsin Inhibitor

HIV protease

Pepstatin

ICE and related enzymes

UP827080, UP827090, UP831910

Proteasome

UP831920, UP831940, UP831960

Thermolysin

Phosphoramidon

Protease inhibitors from Uptima

Ac-Asp-Glu-Val-(Haldehyde)

UP827070

5mg

CA-074

UP827130

5mg

Inhibits apopain/CPP32/Yama

M.Enari et al., 1996, Nature, 376-337

Inhibits cathepsin B

T. Inubushi et al., 1994, J. Biochem, 116:282
Ac-Tyr-Val-Ala-Asp-H(aldehyde) UP827080 5mg CA-074 Me UP827180

5mg

Inhibits ICE and related enzymes

M. Enari et al., 1996, Nature, 380:723.

Inhibits intracellular cathepsin B

D. Buttle et al., 1992, Arch. Biochem. Biophys., 299:377
Ac-Tyr-Val-Lys-Asp-H(aldehyde) UP827090 5mg Chymostatin UP29706B 5mg

Inhibits ICE and related enzymes

T. Graybill et al., 1994, Int. J. Pept. Protein Res., 44:173

MW 605

Inhibits a-,ß-,g-,d-Chymotrypsin, and also many other cysteine proteases (papain)

Working range is 6 - 60 µg/ml (10 - 100 µM)
AEBSF UP40107 E-64 UP789581 5mg

MW 357.4

Inhibits irreversibly cysteine proteases (papain, calpain, cathepsin B,H,L,S)

Working range is 0.5 - 10 µg/ml (0,14 - 28 µM)

Antipain HCl UP257317 5mg E-64 -c UP827140 5mg

MW 677.6

Inhibits reversibly cysteine and serine proteases (trypsin, papain, cathepsin A and B)

Working range is 1-100µM (50 µg/ml = 74 µM)

Inhibits thiol proteases (Cathepsin B/H/L, Calpain)

K. Suzuki, 1983, J. Biochem., 93:1305

Aprotinin UP185582 10mg E- 64 -d UP827190 5mg
UP185586

50mg

MW 651.2

Inhibits competitively and reversibly serine proteases (chymotrypsin, trypsin, kallikrein, plasmin).

Working range is 0.06 - 2.0 µg/ml (0.01 - 0.3 µM)

Inhibits thiol proteases (Cathepsin B/H/L, Calpain)

Chem. Pharm. Bull., 35:1098; M. Tamai et al., 1987
Bestatin UP300991 10mg EDTA UP036290

1kg
UP03629A 100g

MW 308.4

Inhibits competitively aminopeptidases, especially aminopeptidase B, leucine aminopeptidase and tripeptide aminopeptidase, but not carboxypeptidases

Working range is 1-40 µM; Soluble in Methanol

MW 372.2

CAS: 6381-92-6

Ethylenediamine tetraccetic acid

Disodium salt

Purity > 99.5

Inhibits reversibly metalloproteases by chelating metal ions cofactors.

Working range is 1-10 µM

Biotinyl-Asp-Glu-Val-Asp-H (ald.) UP827100 1mg EGTA UP10075A 10g

Inhibits Apopain/CPP32/Yama

D. Nicholson et al., 1995, Nature, 376:37

MW 380.4

Inhibits reversibly metalloproteases by chelating calcium ions cofactors.
Elafin (Human) UP831970 20µg Z-Asp-CH2-DCB UP831910 5mg

Inhibits elastase from human skin

Biophys. Res. Commun., 185:967 ; M. Tsunemi et al., 1992, Biochem.

Inhibits ICE and related proteases

T. Mashima et al., 1995, Biochem. Biophys. Res. Commun.,209:907

Leupeptin UP827721 5mg Z-Ileu-Glu(OBut)-Ala-Leu-H UP831920 5mg
UP827724 25mg (aldehyde) (PSI)

MW 463.0

Inhibits reversibly cysteine and serine proteases (trypsin, plasmin,

papain, kallikrein, thrombin, cathepsin A,B)

Working range is 1-100 µM

Inhibits proteasomes

E. Traenckner et al., 1994, EMBO J., 13:5433

Pepstatin UP827752 25mg Z-Leu-Leu-H UP831930 5mg
(aldehyde) 

MW 685.9

Inhibits aspartic acid proteases (pepsin, renin, cathepsin D) and also HIV protease

Useful range is 0.7 µg/ml (1 µM); soluble in DMSO, Methanol

H. Umezawa et al., 1970, J. Antibiotics, 23:259

Inhibits calpains

Working range is > 10-6M
Phosphoramidon UP348115 5mg Z-Leu-Leu-Leu-H UP831940 5mg
(aldehyde) 

MW 543.5

N-(a-Rhamnopyranosyloxyhydroxyphosphinyl)-Leu-Trp

Inhibits collagenase, thermolysin and metalloendoproteinases

Working range is 4 - 330 µg/ml (7 - 570 µM)

Inhibits proteasomes

T. Jensen et al., 1995, Cell, 83:129
PMSF UP147376 5g Z-Leu-Leu-Nva-H UP831960 5mg
UP147374 25g (aldehyde)

MW 174.2

Phenyl Methyl Sulfonyl Fluoride

Inhibits irreversibly serine proteases (trypsin, chymotrypsin, kallikrein, subtilisin, thrombin) and also cysteine protease papain Working range is 17 - 170 µg/ml(0.1 - 1 mM)

Soluble in Ethanol, Methanol

Inhibits proteasomes

V.Palombella et al., 1994, Cell, 78:773

Trypsin inhibitor UP158624 1mg

from Soybean, immobilized

Inhibits trypsin and trypsin-like proteases (chymotrypsin and elastase)

G. Feinsten et al., 1974 - Euro. J. Biochem, 43:569-581

See also:

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