PMID- 10542048
OWN - NLM
STAT- MEDLINE
DCOM- 20000110
LR  - 20190620
IS  - 0014-2956 (Print)
IS  - 0014-2956 (Linking)
VI  - 266
IP  - 1
DP  - 1999 Nov
TI  - Why reversing the sequence of the alpha domain of human metallothionein-2 does
      not change its metal-binding and folding characteristics.
PG  - 33-9
AB  - A novel peptide, the backward reading sequence of human metallothionein-2 alpha
      domain, was synthesized and its chemical and spectroscopic properties analyzed.
      This folded retro-alpha domain was able to bind Cd(II) in identical
      stoichiometries with the chemically synthesized alpha domain of
      metallothionein-2. Nearly identical to the alpha domain, Cd-binding retro-alpha
      domain showed a characteristic ultraviolet absorption spectrum with a shoulder at
      245-250 nm (due to cadmium-thiolate charge transfer), and the absorption shoulder
      was abolished by acidification [suggesting mercaptide bonding between Cd(II) and 
      the cysteine residues]. Similar metal-binding capabilities between alpha domain
      and retro-alpha domain were observed also by pH titration and in the reaction
      with the sulfhydryl reagent 5,5'-dithiobis(2-nitrobenzoic acid). A two-state
      cooperativity of the metal-cluster formation was observed spectroscopically in
      the titration of the retro-alpha domain, indicating that the retro-protein is
      foldable. In contrast to other proteins, our results indicate that the reversion 
      of the amino acid sequence for the alpha domain does not change its foldability
      and metal-binding capacity, suggesting that the order of its sequence is not
      critical to the formation of a critical metal-tetrathiolate nucleus. However, CD 
      spectra of the Cd-binding alpha domain and retro-alpha domain showed that the
      reversal direction of the domain sequence backbone significantly affects the
      formation of structure even when it is foldable.
FAU - Pan, P K
AU  - Pan PK
AD  - Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan,
      China.
FAU - Zheng, Z F
AU  - Zheng ZF
FAU - Lyu, P C
AU  - Lyu PC
FAU - Huang, P C
AU  - Huang PC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Eur J Biochem
JT  - European journal of biochemistry
JID - 0107600
RN  - 0 (Dinitrobenzenes)
RN  - 0 (MT2A protein, human)
RN  - 0 (Peptides)
RN  - 0 (cadmium-binding protein)
RN  - 0 (cadmium-metallothionein complex)
RN  - 0 (metallothionein 2 protein, rat)
RN  - 00BH33GNGH (Cadmium)
RN  - 9038-94-2 (Metallothionein)
RN  - I0559FK639 (2,4-dinitrothiocyanatobenzene)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cadmium/*metabolism
MH  - Circular Dichroism
MH  - Cysteine/metabolism
MH  - Dinitrobenzenes/*metabolism
MH  - Humans
MH  - Metallothionein/*chemistry/metabolism
MH  - Molecular Sequence Data
MH  - Peptides/chemical synthesis/chemistry/metabolism
MH  - Protein Binding
MH  - Protein Structure, Tertiary
MH  - Rats
EDAT- 1999/10/29 00:00
MHDA- 1999/10/29 00:01
CRDT- 1999/10/29 00:00
PHST- 1999/10/29 00:00 [pubmed]
PHST- 1999/10/29 00:01 [medline]
PHST- 1999/10/29 00:00 [entrez]
AID - ejb811 [pii]
AID - 10.1046/j.1432-1327.1999.00811.x [doi]
PST - ppublish
SO  - Eur J Biochem. 1999 Nov;266(1):33-9. doi: 10.1046/j.1432-1327.1999.00811.x.