We have previously used an ultrasonic oscillation method (please see file; Ala Reconsti).

It seems very difficult to me to reconstitute in accordance with the order of Outer region, Trans-membrane region, and Cytoplasmic region at this time.   Other helps by some membrane glycoprotein and/or specific glyco-phospholipid seem to be required.

Further, although mechanism is not yet clear, biotin may be helping to integration of Asialoglycoprotein receptor (Single-pass type II membrane protein; N-terminus on the cytoplasmic side) into the membrane structure (please see file; biotin asialoglycoprotein receptor).

Amended on 02 August 2017

Since this question seems to be an important question to me in the cancer research, I have further retrieved my protein database of liver specimens derived from the new proteomics of the PDMD (protein-direct-microsequencing-deciphering) method (please see file; HepG2 fucoidan).

Normal fetal immortal Hc has (1) Multi-pass membrane protein at 9.8% of cell protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 1.4% of cell protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.78% of cell protein, and (4) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.49% of cell protein.   Total membrane proteins become to 12.5% of cell protein.

Healed HepG2 (by fucoidan) has (1) Multi-pass membrane protein at 10.8% of cell protein, (2) Single-pass type I membrane protein at 6.0% of cell protein, and (3) Single-pass type II membrane protein at 1.1% of cell protein.   Total membrane proteins become to 17.9% of cell protein. 

Hepatoma HepG2 (without fucoidan) has (1) Multi-pass membrane protein at 5.8% of cell protein, (2) Single-pass type I membrarotein at 7.2% of cell protein, and (3) Single-pass type II membrane protein at 0.45% of cell protein.   Total membrane proteins become to 13.4% of cell protein. 

Ratio of Multi-pass  membrane protein seems to be an indicator of normal or non-cancer state of the cell.   Low ratio of single-pass type I membrane protein seems to be a characteristics of normal fetal cells.

Therefore, notorious "fluid mosaic model" (by Drs. Seymour Jonathan Singer and Garth L. Nicolson) seems to be not effective to the study of the membrane structure, since ratio of Multi-pass membrane protein seems to be more important to construct the membrane structure.   Multi-pass membrane protein should be incorporated into the membrane structure other mechanism than of Single-pass membrane protein and anchored-membrane protein. 

Further amended on 3 August 2017

Further, the new proteomics of the PDMD (protein-direct-microsequencing-deciphering) method has been applied onto the liver specimens.

Normal liver tissue (pseudo liver cancer) has (1) Multi-pass membrane protein at 8.8% of tissue protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 7.3% of tissue protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 4.4% of tissue protein, and (4) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.32% of tissue protein.   Total membrane proteins become to 20.8% of tissue protein.

Liver  cirrhosis (LC) tissue (with leprosy) has (1) Multi-pass membrane protein at 8.2% of tissue protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 6.0% of tissue protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 1.2% of tissue protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.39% of tissue protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of tissue protein.   Total membrane proteins become to 15.8% of tissue protein.

Liver cirrhosis (LC) tissue (designated as No.6) has (1) Multi-pass membrane protein at 15.3% of tissue protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 6.5% of tissue protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.11% of tissue protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.23% of tissue protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of tissue protein.   Total membrane proteins become to 22.1% of tissue protein.

Hepatocellular carcinoma (HCC) tissue (designated as No.6) has (1) Multi-pass membrane protein at 1.5% of tissue protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 4.9% of tissue protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.26% of tissue protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of tissue protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.16% of tissue protein.   Total membrane proteins become to 6.9% of tissue protein.

Hepatocellular carcinoma (HCC) tissue (with primary biliary cirrhosis (PBC)) has (1) Multi-pass membrane protein at 6.1% of tissue protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 6.3% of tissue protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.0% of tissue protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.71% of tissue protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of tissue protein.   Total membrane proteins become to 13.1% of tissue protein.

Hepatocellular carcinoma (HCC) tissues have reduced amount of total membrane proteins (median; 13.1%) as compared to non-cancer tissues (median; 19.4%) (p < 0.05; Mann-Whitney's U test, one-tailed test; n1=3, n2=4).    Normal immortal fetal Hc cells also have low total membrane proteins (12.5%).

Both HCV and HIV-1 have membranes or envelopes.   Both HCV and HIV-1 seem to rule the gene expression of membrane proteins (repress the expression of membrane proteins) in order to monopolize the membrane of host cells or in order to keep the host's membrane structure to viral budding.   Therefore, anticancer effect of fucoidan is as follows; i.e., fucoidan (sulphated poly-fucose) first inhibits the retroviral reverse transcriptase (RT) and can reduce the amount of HIV-1 and HCV, and secondly the inhibitory effect on transcription by HCV and HIV-1 on the membrane-protein biosynthesis is reduced, which gives the normal membrane-protein metabolism or homeostasis of membrane protein in host human cells.

I am very grateful to Dr. Marzieh Dehghan (University of Tehran, Department of Bioinformatics, Tehrān, Tehran, Iran) to lead me to these important findings.

Amended on 17 August 2017

Since membrane-protein metabolism seems to be important, I have further studied among human serum (please see file; JMBT alopecia 1). 

Healthy male adult (33 y) has (1) Multi-pass membrane protein at 1.8% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 6.0% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.0% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 1.4% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 9.2% of serum protein.

A girl (8mo with gastritis) has (1) Multi-pass membrane protein at 0.23% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 0.36% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 1.0% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 1.6% of serum protein.   Soluble part of membrane protein is 85%.   Serum thiol-type protease and its activator is present; i.e., Replicase polyprotein 1ab (Human coronavirus 229E; HCoV 229E; 2702-4090) at 17.7 μg/mg of serum protein and Microtubule-associated protein tau (441-758) at 24.5 μg/mg of serum protein.   These protease and protease activator seems to cut the excreted membrane proteins.     

A girl (12mo with common cold; the same person as above) has (1) Multi-pass membrane protein at 0.84% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 1.7% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 1.7% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 4.2% of serum protein.   Soluble part of membrane protein is 0%.   Serum proteases are not present in this serum. 

Adult serum seems to have higher amount of membrane-protein fragments than child serum.    This age-related changes in membrane-protein metabolism suggest that adult and/or old people should take more protein-foodstuffs than younger people.

Biotin deficiency of child;

1y girl (before biotin therapy) has (1) Multi-pass membrane protein at 1.6% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 1.7% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.0% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 3.5% of serum protein.

1y girl (16w of biotin therapy) has (1) Multi-pass membrane protein at 0.0% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 2.1% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 1.3% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 3.4% of serum protein.

1y girl (44w of biotin therapy) has (1) Multi-pass membrane protein at 2.6% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 2.4% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.0% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 5.0% of serum protein.

4mo boy (before biotin therapy) has (1) Multi-pass membrane protein at 1.5% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 0.3% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 1.0% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 2.8% of serum protein.

4mo boy (13w of biotin therapy) has (1) Multi-pass membrane protein at 1.7% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 1.9% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 2.2% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 1.0% of serum protein.   Total membrane proteins become to 6.8% of serum protein.

3y girl (with GSD-1b, before biotin therapy) has (1) Multi-pass membrane protein at 3.9% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 2.2% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.66% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 6.8% of serum protein.

3y girl (with GSD-1b, after biotin therapy and after 2mo of the liver transplantation) has (1) Multi-pass membrane protein at 4.8% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 1.6% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 3.1% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 9.5% of serum protein.

Liver transplantation seems to increase membrane proteins in serum.

Biotin deficiency of adult;

32y female (with Gait disorder, before biotin therapy) has (1) Multi-pass membrane protein at 4.8% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 1.8% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 1.7% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.18% of serum protein.   Total membrane proteins become to 8.5% of serum protein.

22y female (sister of the patient; with light Gait disorder) has (1) Multi-pass membrane protein at 5.0% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 0.8% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 1.4% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 7.2% of serum protein.

52y female (mother of the patient, healthy) has (1) Multi-pass membrane protein at 4.1% of serum protein, (2) Single-pass type I membrane protein (N-terminus on the extracellular side) at 1.9% of serum protein, (3) Single-pass type II membrane protein (N-terminus on the cytoplasmic side) at 0.0% of serum protein, (4) Single-pass type IIIb membrane protein (N-terminus is membrane anchor) at 0.0% of serum protein, and (5) Single-pass type IV membrane protein (C-terminus is membrane anchor) at 0.0% of serum protein.   Total membrane proteins become to 6.0% of serum protein.

Disease of cerebellum seems to increase membrane-protein fragments in the serum, although the relation of prion (this patient unquely has prion in her serum) and inceased excretion of membrane proteins has not been clarified yet.