PMID- 35163231
OWN - NLM
STAT- MEDLINE
DCOM- 20220307
LR  - 20220307
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 23
IP  - 3
DP  - 2022 Jan 24
TI  - Protein Biochemistry and Molecular Modeling of the Intra-Melanosomal Domain of 
      Human Recombinant Tyrp2 Protein and OCA8-Related Mutant Variants.
LID - 10.3390/ijms23031305 [doi]
LID - 1305
AB  - Tyrosinase-related protein 2 (Tyrp2) is involved in the melanogenesis pathway, 
      catalyzing the tautomerization of dopachrome to 5,6-dihydroxyindole-2-carboxylic 
      acid (DHICA). Recently, a new type of albinism was discovered with 
      disease-causing mutations in the TYRP2 gene. Here, for the first time, we 
      characterized the intra-melanosomal protein domain of Tyrp2 (residues 1-474) and 
      missense variants C40S and C61W, which mimic the alterations found in genetic 
      studies. Recombinant proteins were produced in the Trichoplusia Ni (Ti. Ni) 
      larvae, purified by a combination of immobilized metal affinity (IMAC) and 
      gel-filtration (GF) chromatography, and biochemically characterized. The mutants 
      showed the protein expression in the lysates such as the wild type; however, 
      undetectable protein yield after two steps of purification exhibited their 
      misfolding and instability. In addition, the misfolding effect of the mutations 
      was confirmed computationally using homology modeling and molecular docking. 
      Together, experiments in vitro and computer simulations indicated the critical 
      role of the Cys-rich domain in the Tyrp2 protein stability. The results are 
      consistent with molecular modeling, global computational mutagenesis, and 
      clinical data, proving the significance of genetic alterations in cysteine 
      residues, which could cause oculocutaneous albinism type 8.
FAU - Dolinska, Monika B
AU  - Dolinska MB
AD  - National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
FAU - Woods, Taariq
AU  - Woods T
AD  - National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
FAU - Osuna, Isabella
AU  - Osuna I
AD  - National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
FAU - Sergeev, Yuri V
AU  - Sergeev YV
AUID- ORCID: 0000-0002-7204-6572
AD  - National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
LA  - eng
GR  - ZIA EY000476-10/EY/NEI NIH HHS/United States
PT  - Journal Article
DEP - 20220124
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - 0 (Melanins)
RN  - 0 (Recombinant Proteins)
RN  - EC 1.- (Oxidoreductases)
RN  - EC 5.3.- (Intramolecular Oxidoreductases)
RN  - EC 5.3.3.12 (dopachrome isomerase)
SB  - IM
MH  - Albinism
MH  - Albinism, Oculocutaneous/genetics
MH  - Catalysis
MH  - Humans
MH  - Intramolecular Oxidoreductases/*genetics/metabolism/*ultrastructure
MH  - Kinetics
MH  - Melanins/biosynthesis
MH  - Melanosomes/metabolism
MH  - Models, Molecular
MH  - Molecular Docking Simulation
MH  - Oxidoreductases/metabolism
MH  - Protein Domains/genetics
MH  - Protein Stability
MH  - Recombinant Proteins/metabolism
PMC - PMC8836267
OTO - NOTNLM
OT  - Cys-rich domain
OT  - OCA8-related mutant variants
OT  - effect of reducer
OT  - intra-melanosomal domain
OT  - protein expression/purification
OT  - protein stability
OT  - tyrosinase-related protein 2
COIS- The authors declare no conflict of interest. The funders had no role in the 
      design of the study; in the collection, analyses, or interpretation of data; in 
      the writing of the manuscript; or in the decision to publish the results.
EDAT- 2022/02/16 06:00
MHDA- 2022/03/08 06:00
PMCR- 2022/01/24
CRDT- 2022/02/15 01:16
PHST- 2021/12/08 00:00 [received]
PHST- 2022/01/14 00:00 [revised]
PHST- 2022/01/20 00:00 [accepted]
PHST- 2022/02/15 01:16 [entrez]
PHST- 2022/02/16 06:00 [pubmed]
PHST- 2022/03/08 06:00 [medline]
PHST- 2022/01/24 00:00 [pmc-release]
AID - ijms23031305 [pii]
AID - ijms-23-01305 [pii]
AID - 10.3390/ijms23031305 [doi]
PST - epublish
SO  - Int J Mol Sci. 2022 Jan 24;23(3):1305. doi: 10.3390/ijms23031305.