mccc1  (Proteintech)

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Sociodemographic, biochemical, enzymatic, genetic and clinical information on 88 patients with MCC deficiency 53 Individuals identified by newborn screening without (n = 36) and with (n = 13) reported symptoms (n = 4 without clinical details)

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques:

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Sociodemographic, biochemical, enzymatic, genetic and clinical information on 88 patients with MCC deficiency 18 individuals identified by selective metabolic screening due to clinical symptoms (n = 17, no clinical details in n = 1)

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques: Computed Tomography, Infection, Mutagenesis

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Sociodemographic, biochemical, enzymatic, genetic and clinical information on 88 patients with MCC deficiency 8 individuals identified by family screening (asymptomatic individuals (n = 3), symptomatic individuals (n = 3), no clinical data (n = 2))

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques:

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Sociodemographic, biochemical, enzymatic, genetic and clinical information on 88 patients with MCC deficiency Mothers identified following the positive newborn screening result of their offspring (n = 9)

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques:

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Overview on 64 MCCC1 mutant alleles and their consequences

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques: Mutagenesis

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Expression of MCCC1 and MCCC2 wildtype and mutant alleles

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques: Expressing, Mutagenesis, Plasmid Preparation

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Western blot analysis of expressed MCCC1 wildtype and mutant proteins. Constructs with MCCC1 wildtype and 3 mutant cDNAs in pTracer vector were transfected into MCCC1 deficient reference cell lines by electroporation and harvested 48 hours later for Western blot analysis. 50 μg of protein were used per lane. The MCCC1 subunit was visualized by a) immunostaining using β-actin (4 μg) as control, or by b) colorimetric reaction after coupling with avidin-alkaline phosphatase. Transfection with an empty vector (vector only) was used as a negative control. For further details see “Methods”.

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques: Western Blot, Mutagenesis, Construct, Plasmid Preparation, Transfection, Electroporation, Immunostaining, Avidin-Biotin Assay, Negative Control

Journal: Orphanet Journal of Rare Diseases

Article Title: 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

doi: 10.1186/1750-1172-7-31

Figure Lengend Snippet: Mapping of missense mutations onto the P. aeruginosa MCC holoenzyme structure. ( A ) Mapping of the novel missense mutations (yellow spheres) onto the P. aeruginosa MCC holoenzyme structure (PDB code 3U9T), showing that they are clustered in the BC domain of MCCC1 (pink) or the CT domain of MCCC2 (green). ( B ) and ( C ) Mutation sites of Asn200, Gly220, Pro224 and Gly475 are located at the interface between two MCCC2 subunits (coloured green and purple). ( D ) Structural environment of the MCCC1 Glu288 and Gly379 mutation sites. All the residues shown in sticks are conserved between the human and P. aeruginosa enzymes. The Glu288-Arg444 ionic interaction is indicated by dashed lines.

Article Snippet: The following mutations were introduced by site-directed mutagenesis into the existing wildtype MCCC1 and MCCC2 pCR Blunt II TOPO vector (Invitrogen, Basel, Switzerland): MCCC1 p.E288G, p.G379D, p.I434M and MCCC2 p.S39F, p.G118del, p.Y146N, p.H282R, p.V434L, p.A456V, p.G475R, p.S523G.

Techniques: Mutagenesis