PMID- 10513607
OWN - NLM
STAT- MEDLINE
DCOM- 19991124
LR  - 20190719
IS  - 0918-6158 (Print)
IS  - 0918-6158 (Linking)
VI  - 22
IP  - 9
DP  - 1999 Sep
TI  - Primary structure of rat spermidine synthase: an example of refining the
      cDNA-derived amino acid sequence.
PG  - 889-95
AB  - The primary structure of rat spermidine synthase having the N-terminal acetylated
      methionine and 98.7% homology with that of the mouse enzyme is presented using a 
      limited amount of the homogeneous enzyme. The study strategy was principally to
      compare the molecular masses of liberated peptides determined by three specific
      cleavage methods with those expected from known cDNA-derived amino acid sequences
      of mouse and human enzymes using matrix-assisted laser desorption/ionization
      time-of-flight mass spectrometry (MALDI TOF-MS). The cleavage methods involved
      two enzymatic methods using lysylendopeptidase and arginylendopeptidase, and a
      chemical method for cleaving at the cysteine residue using
      2-nitro-5-thiocyanobenzoic acid. Their usefulness was clearly demonstrated.
      Column-switching semimicro reversed-phase HPLC, which permits application of the 
      entire reaction mixture, was useful for collecting a small amount of peptides
      containing the N-terminal amino acid, to confirm acetylation of the N-terminal
      methionine by MALDI TOF-MS. It was necessary in this approach to examine the
      amino acid sequence of certain peptides. The Edman method was used for the
      sequence analysis, and this will be replaced by an improved MALDI TOF-MS now
      available in a few laboratories.
FAU - Wada, M
AU  - Wada M
AD  - Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Josai
      University, Sakado, Saitama, Japan.
FAU - Amano, D
AU  - Amano D
FAU - Hosoda, H
AU  - Hosoda H
FAU - Shirahata, A
AU  - Shirahata A
FAU - Samejima, K
AU  - Samejima K
FAU - Pegg, A E
AU  - Pegg AE
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Japan
TA  - Biol Pharm Bull
JT  - Biological & pharmaceutical bulletin
JID - 9311984
RN  - 0 (DNA, Complementary)
RN  - 0 (Thiocyanates)
RN  - EC 2.5.1.16 (Spermidine Synthase)
RN  - EC 3.4.- (Endopeptidases)
RN  - O5VCA1GS5O (2-nitro-5-thiocyanobenzoic acid)
SB  - IM
MH  - Acetylation
MH  - Amino Acid Sequence
MH  - Animals
MH  - DNA, Complementary/genetics
MH  - Endopeptidases/metabolism
MH  - Humans
MH  - Male
MH  - Mice
MH  - Molecular Sequence Data
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Sequence Analysis, Protein
MH  - Sequence Homology, Amino Acid
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
MH  - Spermidine Synthase/*chemistry/genetics/metabolism
MH  - Thiocyanates/chemistry
EDAT- 1999/10/08 00:00
MHDA- 1999/10/08 00:01
CRDT- 1999/10/08 00:00
PHST- 1999/10/08 00:00 [pubmed]
PHST- 1999/10/08 00:01 [medline]
PHST- 1999/10/08 00:00 [entrez]
AID - 10.1248/bpb.22.889 [doi]
PST - ppublish
SO  - Biol Pharm Bull. 1999 Sep;22(9):889-95. doi: 10.1248/bpb.22.889.