The use of maleic anhydride for the reversible blocking of amino groups in polypeptide chains.
1. Maleic anhydride was shown to react rapidly and specifically with amino groups of proteins and peptides. Complete substitution of chymotrypsinogen was achieved under mild conditions and the extent of reaction could be readily determined from the spectrum of the maleyl-protein. 2. Maleyl-proteins are generally soluble and disaggregated at neutral pH. Trypsin splits the blocked proteins only at arginine residues and there is frequently selectivity in this cleavage, e.g. in yeast alcohol dehydrogenase and pig glyceraldehyde 3-phosphate dehydrogenase. 3. The group is removed by intramolecular catalysis at acid pH. The half-time was 11-12hr. at 37 degrees at pH3.5 in in-maleyl-lysine or in maleyl-chymotrypsinogen. 4. The unblocking reaction can be used as the basis for a ;diagonal'-electrophoretic separation of lysine peptides and N-terminal peptides, as shown by studies with beta-melanocyte-stimulating hormone.
D000429:Alcohol Oxidoreductases; D000596:Amino Acids; D000812:Anhydrides; D001120:Arginine; D002247:Carbon Isotopes; D055598:Chemical Phenomena; D002621:Chemistry; D002918:Chymotrypsin; D004355:Drug Stability; D004586:Electrophoresis; D004792:Enzyme Precursors; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D006863:Hydrogen-Ion Concentration; D008239:Lysine; D008293:Malates; D009074:Melanocyte-Stimulating Hormones; D008956:Models, Chemical; D010455:Peptides; D011506:Proteins; D012995:Solubility; D013053:Spectrophotometry; D013997:Time Factors; D014357:Trypsin
D000596:Amino Acids; D000812:Anhydrides; D002247:Carbon Isotopes; D004792:Enzyme Precursors; D008293:Malates; D010455:Peptides; D011506:Proteins; D009074:Melanocyte-Stimulating Hormones; D001120:Arginine; D000429:Alcohol Oxidoreductases; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D002918:Chymotrypsin; D014357:Trypsin; D008239:Lysine
Biochem J. 1955 Aug;60(4):541-56
13249947
Biochemistry. 1968 May;7(5):1941-50
5689844
Biochem J. 1968 Sep;109(2):312-4
5679376
Eur J Biochem. 1967 Jul;2(1):84-9
4866857
Nature. 1967 Dec 23;216(5121):1181-5
4294736
Hoppe Seylers Z Physiol Chem. 1957;309(4-6):219-20
13513014
Biochem J. 1959 Mar;71(3):434-45
13638248
Hoppe Seylers Z Physiol Chem. 1968 Feb;349(2):265
5677013
Biochem J. 1966 Oct;101(1):214-28
5971783
Biochem J. 1952 Sep;52(1):3-14
13018153
Biochem J. 1948;42(2):287-94
16748281
Virology. 1965 Aug;26(4):558-66
5833314
Biochemistry. 1963 May-Jun;2:445-52
14069526
Biochim Biophys Acta. 1965 Dec 16;111(2):551-2
5879490
Nature. 1966 Aug 6;211(5049):591-3
5968723
Biochim Biophys Acta. 1958 Sep;29(3):587-93
13584362
Biochem J. 1967 Jun;103(3):78P-79P
6049401
Biochim Biophys Acta. 1952 Dec;9(6):704-5
13032186
Nature. 1968 Sep 7;219(5158):1025-8
4299800
J Biol Chem. 1967 Oct 25;242(20):4752-8
4294024
Biochem J. 1968 Jun;108(2):281-8
4874971
Biochemistry. 1963 Jan-Feb;2:104-16
14001462