MCM6 Antibodies

DNA replication licensing factor MCM6 (MCM6)

Acts as component of the MCM2-7 complex (MCM complex) which is the putative replicative helicase essential for 'once per cell cycle' DNA replication initiation and elongation in eukaryotic cells. The active ATPase sites in the MCM2-7 ring are formed through the interaction surfaces of two neighboring subunits such that a critical structure of a conserved arginine finger motif is provided in trans relative to the ATP-binding site of the Walker A box of the adjacent subunit.

The six ATPase active sites, however, are likely to contribute differentially to the complex helicase activity. .

Gene Information

Human
Gene Name:	MCM6
Uniprot:	Q14566
Entrez:	4175

Family

Belongs to:
MCM family

Alternative Names

DNA replication licensing factor MCM6; EC 3.6.4.12; MCG40308; MCM6 minichromosome maintenance deficient 6 (MIS5 homolog, S. pombe) (S.cerevisiae); MCM6 minichromosome maintenance deficient 6 (MIS5 homolog, S. pombe); minichromosome maintenance complex component 6; minichromosome maintenance deficient (mis5, S. pombe) 6; minichromosome maintenance deficient 6 homolog (S. cerevisiae); minichromosome maintenance deficient 6 homolog; MIS5 homolog; Mis5; p105MCM

Molecular Weight

Mass (kDA):
92.889 kDA

Genomic Context

Human
Location:	2q21.3
Sequence:	2; NC_000002.12 (135839626..135876443, complement)

Subcellular Localizations

Nucleus. Binds to chromatin during G1 and detach from it during S phase.

Diseases

• Neoplasms
• Malignant Neoplasms
• Lactose Intolerance
• Retinoblastoma
• Carcinoma
• Cell Transformation, Neoplastic
• Lung Neoplasms
• Malignant Paraganglionic Neoplasm
• Malignant Neoplasm Of Lung
• Melanoma
• Non-small Cell Lung Carcinoma
• Dysplasia
• Lymphoma

• Adenocarcinoma
• Epithelioma
• Craniopharyngioma
• Vascular Diseases
• Hyperplasia
• Papillary Neoplasm
• Genomic Instability

Interacting Proteins

• CDKN2A
• CDT1
• FAM161A
• MCM10
• MCM2

• MCM3
• MCM7
• MCMBP
• NUDT2
• SNRPB2

• SSRP1
• ZBTB9

Pathways

• Dna Replication
• Cell Cycle
• S Phase
• Localization
• Cell Proliferation
• Cell Division
• Mitosis
• G1 Phase
• Cell Growth
• Dna Replication Initiation
• Transport
• Cell Cycle Arrest
• Regulation Of Dna Replication

• Interphase
• Dna Repair
• Pathogenesis
• Translation
• Cell Death
• Single-stranded Dna Binding
• M Phase

PTMs

• Phosphorylation
• Cleavage

• Carboxylation
• Methylation

• Dephosphorylation

Research Areas

• Cell Cycle and Replication
• DNA Repair

For details, visit:
bosterbio.com/bosterbio-gene-info-cards/MCM6

Best Uses Of The MCM6 Marker

Steven Boster was the inventor of his first products in 1993. This earned him the nickname "he that converts science in a lavatory." His company quickly became China’s leading catalog antibody manufacturer. PicoKine(tm), which he also invented, is a proprietary ELISA system. The company has proprietary trade secrets and develops high-sensitivity ELISA kits for a variety of applications.

Histology

MCM6 and Ki-67 are two clinically useful markers in breast cancer. MCM6 and Ki-67 correlate with mitotic numbers, but MCM6 is significantly more common in tumors. MCM6 can differentiate between different histological grade of invasive-ductal carcinoma. MCM6 is however more specific. Ki-67 has a stronger association than MCM6 with ER status or HR negative tumors.

Numerous types and forms of cancer are also linked to the MCM6 mutation. It has been shown that it is more prevalent in certain types of cancers, such as colon cancer, endometrial cancer and cervical cancer. The marker may also be a prognostic indicator for the progression of OC. The molecular mechanism that underlies the MCM Complex is not yet understood. The MCM family members are promising targets for anti-cancer drugs development.

IHC staining showed that MCM6 was present in tumor tissues. The expression levels were significantly higher than in normal tissue. Furthermore, high expression of MCM6 in HCC tissue was associated with a poorer prognosis. Further, MCM6 is believed to be a possible oncogene in HCC. To establish its clinical significance in this disease, further research is needed.

Despite its importance, little is known about its role in metabolic pathways. However, several types of cancer have been linked to the MCM complex. MCM6 has been implicated in immune progression dysfunction. This protein may be a target for immunotherapy against glioma and systemic lupus erythematosus. MCM6 was also implicated in the development and testing of immunotherapies against anaplastic Oligodendroglioma.

MCM6 has been linked to CDK5RAP3. MCM6 staining in gastric cancer is primarily in the nucleus. MCM6 staining in adjacent non-tumor tissue was found in cytoplasm. This finding shows that MCM6 and CDK5RAP3 expression levels were not significantly different between non-tumor and tumor tissues. However, it does suggest the potential role for MCM6 in predicting poor prognosis in gastric carcinoma.

Primary antibodies with high affinity

The MCM6 protein is a nuclear location marker for chromatin. This marker is localized at discrete foci on spread nucleoids. To detect SpMcm6p we used an antibody that was affinity purified against SpMcm6. We observed SpMcm6p signals in the nuclei of cdc10-129 mutant cells and in HU-cultured early-S arrested wild-type cells.

The MCM6 protein has 821 amino acid residues and two conserved amino motifs. A synthetic peptide that contained amino acid positions 9 to 26 was used for the production of antibodies. The proteins were separated by electrophoresis using denaturing Polyacrylamide gel electrophoresis. After that, the antibodies were blotted with indicated antibodies. Protein extracts of HeLa cells were analyzed with antibodies directed against the MCM6p marker, a protein with an electrophoretic mass of 105 kDa.

High-affinity primary antibodies were used to detect SpMcm6 using BL21 (DE3) transformants. They were grown at 37degC in LB broth containing 60mg/ml methicillin. SDS–PAGE and centriprep concentrators were used for the purification of peptides taken from inclusion bodies. Finally, the SpMcm6 peptide was affinity purified using N-hydroxysuccinimide-activated columns.

This study also revealed the uniqueness of Mcm proteins. As a result, the Mcm proteins are more sensitive to micrococcal nuclease treatment than conventional methods. These antibodies also recognize DNA fragments that contain DEFD-peptides. A similar antibody to MCM6 may be used to detect Mcm proteins or Mcm3p. They also showed good cross-reactivity in chromatin gels, so this method has a wide range of applications.

The MCM6 antibody's pattern is bimodal in both G1 and S phases. During S phase, signal decreases and returns to baseline. The pattern corresponding to the Mcm6 marker shows similar bimodality as Mcm6. The MCM2 antibody, however, has a higher specificity. It is also possible for the MCM6 to detect non-specific antibody.

However, this analysis still leaves some uncertainty in the G1/S interface. Despite the positive immunoreactivity for Mcm proteins, this study also revealed that binding to PCNA improved the marker's ability distinguish G1 cells from S phases. The Mcm6 binding antibody also showed a correlation with BrdU incorporation. These experiments, regardless of cell type, are a good starting point for developing new antibodies to detect Mcm proteins.

UpSetR

The MCM6 marker can detect silent mutations in cells. This is one of its many strengths. The SoupX package provided the "QuickMarkers function" to analyze this marker. This package also allowed for the analysis of the second most hit marker. The MCM6 markers analyzed the data in two groups. One group was created for each IgLV and one group for J genes.