Human G protein alpha subunit gail with at least one mutated amino acid residue

Patent metadata

Field	Value
Publication number	US-10899805-B2
Application number	US-201515516458-A
Country	US
Kind code	B2
Filing date	Sep 2, 2015
Priority date	Oct 2, 2014
Publication date	Jan 26, 2021
Grant date	Jan 26, 2021

Abstract

The present application discloses in detail mutant ligands of the human G protein alpha-subunit -Gαi1-, wherein at least one amino acid residue has been replaced with alanine if the at least one amino acid residue is a non-alanine residue, or at least one amino acid residue has been replace with glycine if the at least one amino acid residue is alanine and wherein the at least one amino acid residue is comprised in a first group containing of the amino acid residues with position R32A, K54A, I55A, I56A, H57A, R176A, E245A, Y296A, T327A, N331A, V332A and D350A or is comprised in a second group containing G42A, A59G, T177A, D200A, A226G, E297A, A300G and F334A or is comprised in a third group containing V50A, A59G, R178A 30 and K180A.

First claim

The invention claimed is:

1. A mutant human G protein alpha-subunit -Gα i1 -, wherein:
(A) at least one amino acid residue of wild-type of the human G-protein-alpha-subunit-Gα i1 -, has been replaced;
(B) the at least one amino acid residue replaced is selected from (1) a first group consisting of the amino acid residues with position R32A, K51A, K54A, I55A, I56A, H57A, R176A, E245A, T327A, N331A, V332A, and D350A (2) a second group consisting of amino acid residues with position G42A, A59G, T177A, D200A, A226G, E297A, A300G, and F334A, or (3) a third group consisting of amino acid residues with position V50A, R178A, and K180A; and
(C) the wild-type of the human G protein alpha-subunit -Gα i1 - comprises residues 23-380 of SEQ ID NO:1:
MGCTLSAEDKAAVERSKMIDRNLREDGEKAAREVKLLLLGAGESGKSTIVKQMKIIHEAGYSEE ECKQYKAVVYSNTIQSIIAIIRAMGRLKIDFGDSARADDARQL VLAGAAEEGFMTAELAGVI KRLWKDSGVQACFNRSREYQLNDSAAYYLNDLDRIAQPNYIPTQQDVLRTRVKTTGIVETHFTF KDLHFKMFDVGGQRSERKKWIHCFEGVTAIIFCVALSDYDLVLAEDEEMNRMHESMKLF DSICNNKWFTDTSIILFLNKKDLFEEKIKKKEIYTHFTCATDTKNVQFVFDAVTDVIIKNCGLF.

2. A mutant Gα protein comprising a mutant human G protein alpha-subunit -Gα a1 -, wherein:
(A at least one amino acid residue of wild-type human G protein alpha-subunit-Gα i1 - has been replaced with
(B) the at least one amino acid residue replaced is selected from (1) a first group consisting of R32G.HNS1.3, K51G.H1.6, K54G.H1.9, I55G.H1.10, I56G.H1.11, H57G.H1.12, R176H.HF.6, E245G.H3.4, T327G.S6H5.4, N331G.H5.3, V332G.H5.4, and D350G.H5.22 (2) a second group consisting of G42G.S1H1.3, A59G.H1HA.2, T177G.HFS2.1, D200G.S3.7, A226G.S4.7, E297G.H4.2, A300G.H4.5, and F334G.H5.6, (3) a third group consisting of V50G.H1.5, R178G.HFS2.2, and K180G.HFS2.4,
(C a common G protein numbering system (CGN) is used in the first, second, and third groups, wherein in said common G protein numbering system, the superscript next to the residue number denotes (i) either the GTPase (G) or helical (H) domain, (ii) the secondary structure element within each domain, and (iii) its position within this structural element according to a sequence alignment of 973 G protein sequences And
(D) (the wild-type of the human G protein alpha-subunit -Gα i1 - comprises residues 23-380 of SEQ ID NO:1:
MGCTLSAEDKAAVERSKMIDRNLREDGEKAAREVKLLLLGAGESGKSTIVKQMKIIHEAGYSEE ECKQYKAVVYSNTIQSIIAIIRAMGRLKIDFGDSARADDARQL VLAGAAEEGFMTAELAGVI KRLWKDSGVQACFNRSREYQLNDSAAYYLNDLDRIAQPNYIPTQQDVLRTRVKTTGIVETHFTF KDLHFKMFDVGGQRSERKKWIHCFEGVTAIIFCVALSDYDLVLAEDEEMNRMHESMKLF DSICNNKWFTDTSIILFLNKKDLFEEKIKKKEIYTHFTCATDTKNVQFVFDAVTDVIIKNCGLF.

Assignees

• Scherrer Inst Paul

Inventors

• Schertler Gebhard
• Sun Dawei
• Veprintsev Dmitry

Classifications

• C12N9/00

  Enzymes; Proenzymes; Compositions thereof (preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; {enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113}); Processes for preparing, activating, inhibiting, separating or purifying enzymes (preparation of malt C12C1/00) · CPC title

• G01N2333/726

  G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH · CPC title

• G01N2500/00

  Screening for compounds of potential therapeutic value · CPC title

• C07K1/00

  General methods for the preparation of peptides {, i.e. processes for the organic chemical preparation of peptides or proteins of any length} · CPC title

• C07K14/4722 Primary

  G-proteins · CPC title