CNBr/formic acid reactions of methionine- and trifluoromethionine-containing lambda lysozyme: Probing chemical and positional reactivity and formylation side reactions by mass spectrometry

被引：25

作者：

Duewell, HS ^{[1
]}

Honek, JF ^{[1
]}

机构：

[1] Univ Waterloo, Dept Chem, Waterloo, ON N2L 3G1, Canada

来源：

JOURNAL OF PROTEIN CHEMISTRY | 1998年 / 17卷 / 04期

关键词：

cyanogen bromide reactions; trifluoromethionine; HPLC-mass spectrometry analysis; formylation;

D O I：

10.1023/A:1022555232364

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The cyanogen bromide (CNBr)/formic acid cleavage reactions of wild-type and trifluoromethionine (TFM)-containing recombinant lambda lysozyme were studied utilizing ESI and MALDI mass spectrometry. Detailed analysis of the mass spectra of reverse-phase HPLC-purified cleavage fragments produced from treatment of the wild-type and labeled proteins with CNBr indicated cleavage solely of methionyl peptide bonds with no observation of cleavage at TFM. N-Acetyl-TFM was also found to be resistant to reaction with CNBr, in contrast to N-acetyl-methionine. The analysis also indicated differential reactivity among the three methionine positions in the wild-type enzyme. Additionally, formylation of intact enzyme as well as peptide fragments were observed and characterized and indicated that serine, threonine, as well as C-terminal homoserine side chains are partially formylated under standard cleavage protocols.

引用

页码：337 / 350

页数：14

共 29 条

[1] PROCESSING OF THE INITIATION METHIONINE FROM PROTEINS - PROPERTIES OF THE ESCHERICHIA-COLI METHIONINE AMINOPEPTIDASE AND ITS GENE STRUCTURE [J].

BENBASSAT, A ;

BAUER, K ;

CHANG, SY ;

MYAMBO, K ;

BOOSMAN, A ;

CHANG, S .

JOURNAL OF BACTERIOLOGY, 1987, 169 (02) :751-757

[2] CYANOGEN-BROMIDE TREATMENT OF N-ACETYLMETHIONYL RESIDUES WITHOUT CLEAVAGE [J].

CARPENTER, FH ;

SHIIGI, SM .

BIOCHEMISTRY, 1974, 13 (25) :5159-5164

[3]

Coligan J. E., 1996, CURRENT PROTOCOLS PR

[4] Use of F-19 NMR to probe protein structure and conformational changes [J].

Danielson, MA ;

Falke, JJ .

ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 1996, 25 :163-195

[5]

DELL A, 1986, PRACTICAL PROTEIN CH, P531

[6] PARTIAL NON-CLEAVAGE BY CYANOGEN-BROMIDE OF A METHIONINE-CYSTINE BOND FROM HUMAN-SERUM ALBUMIN AND BOVINE ALPHA-LACTALBUMIN [J].

DOYEN, N ;

LAPRESLE, C .

BIOCHEMICAL JOURNAL, 1979, 177 (01) :251-254

[7] Incorporation of trifluoromethionine into a phage lysozyme: Implications and a new marker for use in protein F-19 NMR [J].

Duewel, H ;

Daub, E ;

Robinson, V ;

Honek, JF .

BIOCHEMISTRY, 1997, 36 (11) :3404-3416

[8] INVESTIGATIONS OF THE INTERACTIONS OF SACCHARIDES WITH THE LYSOZYME FROM BACTERIOPHAGE-LAMBDA [J].

DUEWEL, HS ;

DAUB, E ;

HONEK, JF .

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1995, 1247 (01) :149-158

[9] F-19 nuclear magnetic resonance chemical shifts of fluorine containing aliphatic amino acids in proteins: Studies on Lactobacillus casei dihydrofolate reductase containing (2S,4S)-5-fluoroleucine [J].

Feeney, J ;

McCormick, JE ;

Bauer, CJ ;

Birdsall, B ;

Moody, CM ;

Starkmann, BA ;

Young, DW ;

Francis, P ;

Havlin, RH ;

Arnold, WD ;

Oldfield, E .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1996, 118 (36) :8700-8706

[10]

Fontana A., 1986, PRACTICAL PROTEIN CH, P67

← 1 2 3 →