Glutaredoxin from rabbit bone marrow. Purification, characterization, and amino acid sequence determined by tandem mass spectrometry.
A glutaredoxin was purified from rabbit bone marrow, and its amino acid sequence was determined by high performance tandem mass spectrometry. The sequences of peptides generated by digestion with trypsin alone or in combination with thermolysin were determined from their collision-induced dissociation (CID) mass spectra. Alignment of these sequences and additional sequence information were obtained from the collision-induced dissociation mass spectra of peptides obtained from digestion of the intact protein with Staphylococcus aureus V8 protease and alpha-chymotrypsin. The resulting sequence of 106 amino acids is as follows: Ac-Ala-Gln-Glu-Phe-Val-Asn-Ser-Lys-Ile-Gln-Pro-Gly-Lys-Val-Val-Val-Phe- Ile-Lys-Pro-Thr-Cys-Pro-Tyr-Cys-Arg-Lys-Thr-Gln-Glu-Ile-Leu-Ser-Glu-Leu- Pro-Phe - Lys-Gln-Gly-Leu-Leu-Glu-Phe- Val-Asp-Ile-Thr-Ala-Thr-Ser-Asp-Met-Ser-Glu-Ile- Gln-Asp-Tyr-Leu-Gln-Gln-Leu-Thr-Gly-Ala-Arg- Thr-Val-Pro-Arg-Val-Phe-Leu-Gly-Lys-Asp-Cys-Ile- Gly-Gly-Cys-Ser-Asp-Leu-Ile-Ala-Met-Gln-Glu-Lys- Gly-Glu-Leu-Leu-Ala-Arg-Leu-Lys-Glu-Met-Gly- Ala-Leu-Arg-Gln. This glutaredoxin strongly resembles the corresponding calf and pig proteins (known as glutaredoxin and thioltransferase, respectively) with respect to its primary structure and enzymatic activity as a GSH:disulfide thioltransferase, an activity also found for the glutaredoxin from Escherichia coli. However, rabbit glutaredoxin was not active as a hydrogen donor for the reduction of ribonucleotides in the presence of the ribonucleotide reductases from rabbit bone marrow, Lactobacillus leichmannii, and Corynebacterium nephridii.
Hopper
S
S
Department of Pharmacology/Physiology, School of Dental Medicine, University of Pittsburgh, Pennsylvania 15261.
Johnson
R S
RS
Vath
J E
JE
Biemann
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001853:Bone Marrow / Q000032:analysis*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000032:analysis; D054477:Glutaredoxins; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010088:Oxidoreductases; D010446:Peptide Fragments / Q000302:isolation & purification; D010447:Peptide Hydrolases; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification; D011817:Rabbits; D012689:Sequence Homology, Nucleic Acid; D013950:Thymus Gland / Q000032:analysis; D014357:Trypsin
D001426:Bacterial Proteins; D054477:Glutaredoxins; D010446:Peptide Fragments; D011506:Proteins; D010088:Oxidoreductases; D010447:Peptide Hydrolases; D014357:Trypsin