PXD034893

PXD034893 is an original dataset announced via ProteomeXchange.

Title	Protein expression and identification of Tyrp1 Mutant Variants Associated with OCA3 form of albinism.
Description	Oculocutaneous albinism type 3 (OCA3) is an autosomal recessive disorder caused by mutations in the TYRP1 gene. Tyrosinase-related protein 1 (Tyrp1) is involved in eumelanin synthesis, catalyzing the oxidation of 5,6-dihydroxyindole-2-carboxylic acid oxidase (DHICA) to 5,6-indolequinone-2-carboxylic acid (IQCA). In our work, we are trying to understand the effect of genetic mutations at the level of protein atomic structure and establish a link between the effect of mutations and disease phenotype. Here, for the first time, four OCA3-causing mutations of Tyrp1, C30R, H215Y, D308N, and R326H, were investigated computationally and experimentally to understand Tyrp1 protein stability and catalytic activity. Using the Tyrp1 crystal structure (PDB:5M8L), global mutagenesis was conducted to evaluate mutant protein stability. Consistent with our predictions the foldability parameter, C30R and H215Y should exhibit greater instability, and two other mutants, D308N and R326H, are expected to keep a native conformation. Mass-spectroscopy analysis has confirmed the identity of mutant recombinant variants. SDS-PAGE and Western blot analysis of the purified recombinant proteins confirmed that the foldability parameter correctly predicted the effect of mutations critical for protein stability. C18 data see C4 data.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:00:39.258.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD034893
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	David Anderson
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2022-06-26 07:22:53	ID requested
1	2023-06-12 04:57:59	announced
⏵ 2	2023-11-14 09:00:41	announced	2023-11-14: Updated project metadata.

Dolinska MB, Anderson DE, Sergeev YV, In vitro characterization of the intramelanosomal domain of human recombinant TYRP1 and its oculocutaneous albinism type 3-related mutant variants. Protein Sci, 32(1):e4518(2023) [pubmed]

10.6019/PXD034893;

submitter keyword: OCA3,Albinism, Tyrp1

Brian P. Brooks
contact affiliation	Pediatric, Developmental & Genetic Ophthalmology Section AND Ophthalmis Clinical Genetics Section, Ophthalmic Genetics and Visual Function Branch, National Eye Institute Intramural Research Program, NIH, US Government
contact email	brooksb@mail.nih.gov
lab head
David Anderson
contact affiliation	NIH/NIDDK
contact email	davida@mail.nih.gov
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD034893
     1. Label: PRIDE project
     2. Name: Protein expression and identification of Tyrp1 Mutant Variants Associated with OCA3 form of albinism.