Protein Folding of A Biopharmaceutical — hCD83

Lin Zhang

Chemical Engineering

Department

Biopharmaceuticals

Proteins

In use: Insulin tPA

Potential use: Human CD83

Outlines

Backgrounds of hCD83

Preliminary Results and Discussion

Human CD83 (hCD83)

Human CD83 is expressed predominantly on the surface of dendritic cells (DCs)

DCs are the most potent antigen presenting cells of the immune system

Glycoprotein CD83 is one of the best-known maturation markers for human DCs

Backgrounds

hCD83

ArgosDC Bio

Production

Preclinical Trial

Backgrounds

Insulin

Backgrounds

What we are interested in:

Dynamic process of protein folding Protein Aggregation 3D Structure

Backgrounds

FDA Approval

3D Structure

Crystallography

3D Structure Prediction Flowchart

Target Sequence

Homologous in PDB

Secondary Structure

Comparative Modeling

Tertiary Structure

Conserved Domain

Yes No

ⅡⅠ

Comparison

Preliminary Results

Database Searching Protein-Protein Blast

Descriptions

Preliminary Results

Preliminary Results

1MCP_H

Class: All beta proteins

Fold: Immunoglobulin-like beta-sandwich

sandwich; 7 strands in 2 sheets; greek-key some members of the fold have additional strands

Superfamily: Immunoglobulin (IG)

Family: V set domains (antibody variable domain-like)

1GL4_B

Class: All beta proteins

Fold: Immunoglobulin-like beta-sandwich

sandwich; 7 strands in 2 sheets; greek-key some members of the fold have additional strands

Superfamily: Immunoglobulin (IG)

Family: I set domains

Preliminary Results

Protein fold recognition Phyre

1nez_g

Preliminary Results

Ⅰ Comparative Modeling

Swiss-Model

RAPTOR

Preliminary Results

1MCP_H

(7~121)

Swiss-Model

Preliminary Results

Swiss-Model

1a6w_L

Discussion

Common

Discussion

Differences

1MCP_H

1α-helix e f

Discussion

Differences

1a6w_L

b c

2α-helices

e f

Preliminary Results

RAPTOR

1a49_a

Preliminary Results

1a49_a

1 α-helix: e f : Thr(83)~Ser(87)

Preliminary Results

Ⅱ Secondary Structure and Tertiary Structure

Secondary Structure: PHD and Jnet

Tertiary Structure : HMMSTR

Preliminary Results

PHD

Preliminary Results

Jnet

Discussion

Secondary Structure

PHD: 6 β-strands, no α-helix, not to be globular proteinV(8)~V(10), D(17)~C(20), V(32)~K(36), S(76)~N(81),

T(89)~L(94), V(107)~T(112)

Jnet: 8 β-strands, no α-helixV(8)~C(12), D(15)~T(21), T(31)~K(36), E(43)~T(47)

N(64)~D(68), Y(75)~N(81), T(89)~Q(95), G(104)~T(112)

Preliminary Results

HMMSTR

Discussion

HMMSTR

3 α- helices:

Asp(98)~Asn(102)

Pro(115)~Arg (118)

Lys(119)~Ile(130)

Discussion

Comparison

A. Fold PatternComparative Modeling: distinguished hydrophobic core and hydrophilic side, regular, tight

HMMSTR: irregular, loose pattern, no related motif

Discussion

Comparison

B. CompositionComparative Modeling: more beta-strands and loops

as well as hydrogen bonds

HMMSTR: less beta-strands, loops and hydrogen bonds