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Enzyme Inhibitors [Biochemicals and Reagents]

 In this section, proteases are categorized into three modes of actions: (1) serine protease; (2) cysteine protease; (3) metalloprotease.1)
Serine proteases are inhibited by AEBSF, benzamidine, etc.2) Cysteine proteases are inhibited by compounds which react with SH groups such as 2-iodoacetamide or 2-iodoacetic acid.3) Metalloproteases are inhibited by chelating agents, EDTA or 1,10-phenanthroline.4)
 In the course of protein extraction, proteolysis is considered to be a major problem because it leads to decreasing yields. Addition of protease inhibitors helps to avoid the proteolysis and improves recovery of the desired protein.5) In the experiment of immunoprecipitation, protease inhibitors are also used to avoid decomposition of antigens or antibodies by proteolytic impurities.6) Protease inhibitors which are frequently used in biochemical research are illustrated in this section.
 Thiol protease inhibitors, 2-iodoacetamide and 2-iodoacetic acid, cannot be used in protein extraction, because they form covalent bonds to the SH group of the proteins. In protein research, they are used as an alkylation reagent to derivatize the proteins which are extracted from an electrophoresis gel for mass spectrometric analysis.7)

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  AEBSF and 2-iodoacetamide gradually decompose in an aqueous solution. Their solutions should be prepared just prior to use.
A0312
6-Aminohexanoic Acid
A0408
3-Aminopropionitrile (stabilized with K2CO3)
A0410
Aminopterin Hydrate
A0630
3-Aminobenzamide
A1163
Procaine Hydrochloride
A1281
3-Aminophenylboronic Acid Monohydrate (contains varying amounts of Anhydride)
A1514
Apigenin
A2033
5-Azacytidine
A2255
6-Aminohexanoic Acid [for Biochemical Research]
A2353
Amlodipine Besylate
A2370
Anastrozole
A2408
W-12 Hydrochloride
A2410
W-5 Hydrochloride
A2416
W-13 Hydrochloride
A2425
Amezinium Methyl Sulfate
A2485
Acarbose Hydrate
A2501
Acetyldinaline
A2528
Acadesine
A2529
PD 98059
A2548
SP 600125
A2598
Acetazolamide
A2599
Amiloride Hydrochloride Hydrate
A2613
Anisomycin
A3351
Salirasib
A5014
TMPyP [=α,β,γ,δ-Tetrakis(1-methylpyridinium-4-yl)porphyrin p-Toluenesulfonate] [Ultra-high sensitive spectrophotometric reagent for Cu, Mg] [For the simultaneous determination of metals by HPLC]
B0099
Benzoyl-β-alanine
B0479
Disulfiram
B0683
Diacetyl Monoxime
B1062
Benzylsulfonyl Fluoride
B1701
PBN
B1890
(+)-Bicuculline
B3082
N-Benzyl-p-toluenesulfonamide
B3803
Butein
B3972
Bleomycin Sulfate (mixture)
B4000
Ubenimex Hydrochloride [for Biochemical Research]
B4006
BIO
B4108
Benserazide Hydrochloride
B4258
Brinzolamide
B4436
TDZD-8
B4823
GO-Y030
B4826
Bikinin
B4845
BAY 11-7085
B4945
PD 153035 Hydrochloride
B5873
N-Butylthiophosphoric Triamide
C0002
Caffeic Acid
C0019
Cantharidin
C0181
Chlorogenic Acid Hydrate
C0187
Clioquinol
C0380
Colchicine (contains 5% Ethyl Acetate at maximum)
C0434
Curcumin (Natural)
C1254
Chlorhexidine Dihydrochloride
C1495
(S)-(+)-Camptothecin
C2243
Gimeracil
C2302
Curcumin (Synthetic)
C2450
Carbidopa Monohydrate
C2481
Chlorpromazine Hydrochloride
C2511
Chlorhexidine Diacetate
C2520
Combretastatin A4
C2663
Carmofur
C2745
Cetilistat
C2867
Clotrimazole
C3009
Combretastatin A4 Phosphate Disodium Salt
C3038
Cloperastine Hydrochloride
C3105
Chlorhexidine Digluconate (20% in Water)
C3183
3-Chloro-5-hydroxybenzoic Acid
D0800
Nordihydroguaiaretic Acid
D0940
2,5-Di-tert-butylhydroquinone
D1817
Methyldopa Sesquihydrate
D2274
Dipyridamole
D2668
Daidzein
D3920
Daidzin
D4015
Daminozide
D4027
Disodium L-(+)-Tartrate Dihydrate
D4061
Diacerein
D4102
Docetaxel
D4126
Doxazosin Mesylate
D4189
Dorzolamide Hydrochloride
D4193
Doxorubicin Hydrochloride
D4292
DRB
D4302
Doxofylline
D4823
PSI-6206
E0136
N-Ethylmaleimide
E0375
Ellagic Acid Dihydrate
E0430
EDTA 3Na Hydrate
E0500
Emodin
E0675
Etoposide
E0694
(-)-Epigallocatechin Gallate Hydrate
E0906
Epalrestat
E1046
BIA 2-093
E1084
(-)-Epigallocatechin
F0675
Finasteride
F0839
Fasudil Hydrochloride
F1007
Fusidic Acid
G0265
(S)-(-)-2-Guanidinoglutaric Acid
G0272
Genistein
G0293
Galantamine Hydrobromide
G0334
Geldanamycin
G0371
Gnetol
H0368
1,2,3,4,5,6-Hexabromocyclohexane
H0409
Hydralazine Hydrochloride
H0912
Haloperidol
H1340
NCC-149
H1388
Vorinostat
H1389
H-Phg(4-OH)-OH
H1525
GO-Y078
H1553
HPA-12
I0044
2-Iodoacetamide
I0804
Isorhapontigenin
I0822
Isoliquiritigenin
I0904
Imperatorin
I0936
Imatinib Mesylate
I0975
PluriSIn 1
K0018
Kaempferol Hydrate
K0052
Kenpaullone
L0214
Lovastatin
L0233
Lansoprazole
L0239
Lornoxicam
L0244
Loxoprofen
L0248
Letrozole
L0252
Loxoprofen Sodium Salt Dihydrate
M0216
Dicoumarol
M1004
4'-Methoxyphenacyl Bromide
M1149
Capsaicin (Natural)
M1663
Milrinone
M1732
Mifepristone
M2131
Myricetin
M2133
Milnacipran Hydrochloride
M2216
Mycophenolic Acid
M2275
Mevastatin
M2302
Miglitol
M2410
LY 294002
M2518
3-Methyladenine
M2530
2-Methoxy-β-estradiol
M2572
URB-754
N0538
6-Nitroveratraldehyde
N0958
NU 2058
N0966
KB-R 7943
O0164
Potassium Oxonate
O0359
Omeprazole
O0373
Oxyresveratrol
O0381
Orlistat
O0400
ODQ
O0450
Oncrasin 1
P0140
Phenylarsine Oxide
P0221
1,10-Phenanthroline Monohydrate
P0823
Valproic Acid
P1632
Paclitaxel
P1796
Pravastatin Sodium
P1923
Polymyxin B Sulfate
P1927
Pinostilbene
P1928
Piceatannol
P1975
Probenecid
P1982
Parthenolide
P2048
Pifithrin-μ
P2088
Phenethyl Caffeate
P2311
Puromycin Dihydrochloride
R0071
Resveratrol
R0087
Risperidone
R0093
Rivastigmine L-Tartrate
R0097
Rapamycin
R0103
Racecadotril
R0115
Rabeprazole Sodium Salt
R0121
Rhododendrol
S0509
Simvastatin
S0519
Sodium Butyrate
S0580
Sulfasalazine
S0868
Sulbactam
S0894
Valproic Acid Sodium Salt
T2353
Niflumic Acid
T2477
Trichostatin A
T2522
H-Cys(Trt)-OH
T2755
Topiramate
T2810
TPCK
T2844
BAPTA-Na4
T2846
BAY 11-7082
T2920
TBB
T2944
Tyrphostin AG 1478
T2962
Tyrphostin AG 490
T2978
Tiropramide Hydrochloride
T3074
Tyrphostin A1
T3101
Talniflumate
T3109
Teniposide
T3732
Tazobactam
V0110
Venlafaxine Hydrochloride
V0119
Voglibose
W0007
Wortmannin
X0077
XAV 939

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References

  • 1)G. G. Guilbault, in Handbook of Enzymatic Methods of Analysis, Clinical and Biochemical Analysis, A series of Monographs and Textbook, ed. by M. K. Schwartz, Marcel Dekker, Inc., New York, 1976.
  • 2)(a) P. Walsmann, M. Richter, F. Markwardt, Acta Biol. Med. Germ. 1972, 28, 577.
    • (b) M. Mares-Guia, E. Shaw, W. Cohen, J. Biol. Chem. 1967, 242, 5777.
  • 3)R. Arnon, Methods Enzymol. 1970, 19, 226.
  • 4)H. Matsubara, Methods Enzymol. 1970, 19, 642.
  • 5)(a) K. Weber, J. R. Pringle, M. Osborn, Methods enzymol. 1972, 26, 3.
    • (b) J. Sambrook, D. W. Russell, in Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Labolatory Press, New York, 2001.
  • 6)P. L. R. Bonner, in Protein Purification, Taylor & Francis, New York, 2007.
  • 7)W. R. Gray, in Protein Function: A Practical Approach, 2nd ed., ed. by T. E. Creighton, Oxford University Press, New York, 1997, Chap. 7, p. 165.
  • 8)Handbook of Inhibitors, ed. by T. Akiyama, K. Kohu, Yodosha, 2006.

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