Hi Fakhriedzwan, lectins are not very specific and they do not usually bind their targets with high affinity.

Another problem with doing work with lectins on live cells is that they mediate intracellular activation and internalization due to "cell capping".

I would recommend using fixed cells for any competition/blocking studies between viruses and lectins.

Dear Stefansson,

Thank you for your input. I'd like to know if viruses are able to bind to non-live cells?

Hi Fakhriedzwan, as far as I know most viruses do not require live cells for the initial cell surface binding because they are binding to a specific target. For example adenovirus binding to integrins.

Live cells are required to internalize the virus.

If fixed cells don't work, then you can do the binding assay at 4oC for ~ 1 hour. At this temperature, the surface proteins are not affected and cell integrity is not compromised. But this low temperature inhibits endocytosis. 

.

Thank you for your response. I really appreciate it

Amended on 26 June 2017

It seems an interesting question for me.   Thus, I have searched Dengue virus (DENV) and gene expression ruled by this virus.   DENV is similar to HCV (Flavivirus, +ssRNA).   HAdV seems to be not linked to integrin.   On the other hand, DENV seems to be linked to Integrin.   Integrin may be also used as a receptor for DENV.  

Normal liver (with pseudo liver cancer) has Genome polyprotein (DENV-4) at 5.2 μg/mg tissue protein, and has expressed Integrin alpha-E/HML-1 antigen/CD103 at 0.16 μg/mg tissue protein and Integrin beta-4 subunit/GP150 at 12.4 μg/mg tissue protein.

LC tissue (designated as No.6) has Genome polyprotein (DENV-4) at 0.31 μg/mg tissue protein, and has expressed Integrin alpha-V/CD51/Vitronectin receptor alpha subunit at 0.38 μg/mg tissue protein.

HCC tissue (designated as No.6) has Genome polyprotein (DENV-4) at 19.8 μg/mg tissue protein, and has expressed Platelet membrane glycoprotein IIIA/CD61/Integrin beta-3/Integrin ITGAV:ITGB3 at 6.1 μg/mg tissue protein.

Then, protein-receptors for virus seem to be ruled and/or induced by virus.   Further, protein-receptors for microbes seem to be membrane glycoproteins.  Therefore, I hope you that your research is successful.

By the way, I would like to know why only DENV-4 is present in Japan. 

Amended on 26 June 2017

Furthermore, I have searched Human adenovirus (HAdV) and gene expression ruled by this virus (dsDNA) based upon new proteomics PDMD (Protein-Direct-Microsequencing-Deciphering) method (please see file; HeoG2 Fucoidan).   Human adenovirus (HAdV) seems to upregulate the expression of GPCR (G protein-coupled receptor; membrane glycoprotein), though its function as a viral receptor or not is not clear at this time.

Cultured hepatoma HepG2 (without fucoidan) has Packaging protein 3 (HAdV-2) at 0.2 μg/mg cell protein, and expresses Taste receptor type 2 member 42/T2R42/T2R55  (GPCR; G protein-coupled receptor) at 0.28 μg/mg cell protein.

Healed normal HepG2 (with fucoidan) has DNA polymerase (HAdV-40) at 0.9 μg/mg cell protein and Fiber protein/Protein IV (HAdV-12) at 0.4 μg/mg cell protein (total 1.3 μg/mg cell protein), and expresses Beta-1 Adrenergic receptor at 0.57 μg/mg cell protein and Leucine-rich repeat-containing G-protein coupled receptor 6 at 0.78 μg/mg cell protein (total 1.35 μg/mg cell protein).

Serum of 1y girl (biotin deficiency) has Maturation protein (HAdV-6) at 7.7 μg/mg serum protein, and expresses Regulator of G-protein signaling 19 at 3.4 μg/mg serum protein.

Normal liver (pseudo liver cancer) has E1A protein (HAdV-7) at 2.0 μg/mg tissue protein, and expresses Mas-related G-protein coupled receptor member X3 at 1.6 μg/mg tissue protein.

LC tissue (with leprosy) has Protein VI (HAdV-2) at 2.9 μg/mg tissue protein, and expresses Mas-related G protein-coupled receptor MRG/MAS-R at 0.88 μg/mg tissue protein.

HCC tissue (with PBC) has Control protein E1A/55-residue (55R) E1A protein (HAdV-2) at 0.08 μg/mg tissue protein, and expresses B2 Bradykinin receptor/BK-2 receptor at 1.2 μg/mg tissue protein.

LC tissue (No.6) has Early E3A 12.5 KD protein (HAdV-2) at 0.5, Hypothetical protein ORF1 (AAdV) at 0.6, and Probable early E4 11KD protein (HAdV-2) at 1.5 μg/mg tissue protein (total of 2.6 μg/mg tissue protein), respectively, and expresses Mas-related G protein-coupled receptor MRG at 5.1 μg/mg tissue protein.

HCC tissue (No.6) has E3B 14.7 KD protein (HAdV-12) at 4.2, Fiber protein (HAdV-12) at 0.31, and 11.6 kD protein (HAdV-2) at 0.06 μg/mg tissue protein, respectively (total of 4.6 μg/mg tissue protein), and expresses Rhodopsin at 2.8 μg/mg tissue protein.

Thus, HAdV may not be directly linked to the cancer, but the use of HAdV and AAdV as a vector in biotechnology may be dangerous due to up-regulation of the G protein-coupled receptors (GPCRs) in the host cells. 

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