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Biosynthesis of Insulin

Donald F. Steiner, Shu Jin Chan, Arthur H. Rubenstein

10.1002/cphy.cp070203

Source: Supplement 21: Handbook of Physiology, The Endocrine System, The Endocrine Pancreas and Regulation of Metabolism

Originally published: 2001

Published online: January 2011

Full Article on Wiley Online Library



Abstract

The sections in this article are:

1 Insulin: Properties and Structure
2 Biosynthesis of Insulin
2.1 Structure and Functions of Precursor Forms
2.2 Cell Biology
2.3 Mechanism of Proteolytic Conversion of Proinsulin to Insulin
2.4 Insulin Storage Vesicles
2.5 C Peptide, a Co‐secretory Product of the β Cell
3 Regulation of Insulin Biosynthesis
4 The Insulin Gene and its Defects
4.1 Mutations in the Insulin Gene
5 Defects in Insulin Biosynthesis
5.1 Prohormone Convertase Defects
6 Conclusion
Figure 1. Figure 1.

Primary structure of human insulin 195 Residues shown in boxes are conserved in all known vertebrate insulins.
Figure 2. Figure 2.

Structure of the two‐zinc porcine insulin hexamer (peptide main chain) based on x‐ray crystallographic data 13. The threefold axis is perpendicular to the page; the two zinc atoms lie on this axis, above and below the center of the hexamer, each coordinated by three B10 histidine side chains. Light and dark ribbons represent dimer pairs shown on their twofold axes (lines).
Figure 3. Figure 3.

Structure of the porcine insulin monomer. The B‐chain peptide backbone is shown as a dark ribbon and the A chain as a light ribbon.
Figure 4. Figure 4.

A: Covalent structure of rat preproinsulin I.

B: Primary sequences of representative vertebrate proinsulins compared with the human sequence. Conserved residues are highlighted. Sources are as follows: human 17, hummingbird 68, Xenopus 265, zebraflsh 179, hagfish 39, amphioxus 38.
Figure 5. Figure 5.

A: Covalent structure of rat preproinsulin I.

B: Primary sequences of representative vertebrate proinsulins compared with the human sequence. Conserved residues are highlighted. Sources are as follows: human 17, hummingbird 68, Xenopus 265, zebraflsh 179, hagfish 39, amphioxus 38.
Figure 6. Figure 6.

Subcellular organization of the insulin biosynthetic pathway. Arrows indicate direction of movement of (pro)insulin via subcellular compartments. Dashed arrow pointing downward indicates the mannose‐6‐P receptor pathway to the lysosome. Dashed arrow pointing upward indicates granule degradative pathway (autophagy). Time scales indicate intracompartmental residence times (see text for details). RER, rough endoplasmic reticulum.
Figure 7. Figure 7.

Schematic diagram showing “constitutive‐like” alternative secretory pathway arising from immature secretory granules (vesicles) via passive sorting of soluble, nonaggregated C peptide generated by proinsulin processing during early phases of secretory granule maturation. Other granule constituents, such as furin and/or lysosomal enzyme precursors, may be actively removed from the maturing granules via clathrin‐clad vesicles by virtue of cytosolic domain interactions (furin) or mannose‐6‐P receptor (M‐6‐PR)–mediated pathways (cathepsins) (see text and ref. 6 for details). TGN, trans‐Golgi network.
Figure 8. Figure 8.

Proinsulin processing pathways in the β cell. On the right is the predominant pathway, beginning with cleavage of proinsulin by prohormone convertase 3 (PC3) to yield des‐31, 32‐proinsulin, a preferred substrate for PC2. However, evidence summarized in the text indicates that either enzyme is capable of cleavage at both junctions in the prohormone to generate insulin. Carboxy peptidase E (CPE) removes C‐terminal basic amino acids from products (not shown) generated by the endoproteolytic action of PC2 or PC3.
Figure 9. Figure 9.

Structures of the subtilisin‐like proprotein convertases. The upper three [Prohormone convertase 2 (PC2), PC1/PC3, and PC4] are neuroendocrine‐specific in their localization and functions, while the lower group (furin, PACE, PC6, and PC7) are expressed more widely and tend to process a variety of constitutively secreted precursors (for review, see ref. 234). Pre, signal peptide; Pro, pro domain; Cat, catalytic domain; P, P domain; CR cysteine‐rich domain; AH, amphipathic helix; TM, transmembrane segment; S/TR, serinethreonine‐rich region. Catalytic residues D, H, N, and S are shown above the catalytic domains. Glycosylation sites are not shown.
Figure 10. Figure 10.

Structure of the insulin gene in selected vertebrates, showing highly conserved intron–exon structure and the promoter region. Numbers indicate variable size (in nucleotides) of each intron.
Figure 11. Figure 11.

Electron micrograph of β cells in wild‐type (PC2 +/+) or mutant (PC‐/‐) mice. Note abundance of mature secretory granules in wild‐type contrasting with large numbers of immature‐appearing secretory granules in the mutant, indicative of increased proinsulin content 78. Bar = 1 mM.

(Photomicrograph courtesy of Hewson H. Swift.)


Figure 1.

Primary structure of human insulin 195 Residues shown in boxes are conserved in all known vertebrate insulins.



Figure 2.

Structure of the two‐zinc porcine insulin hexamer (peptide main chain) based on x‐ray crystallographic data 13. The threefold axis is perpendicular to the page; the two zinc atoms lie on this axis, above and below the center of the hexamer, each coordinated by three B10 histidine side chains. Light and dark ribbons represent dimer pairs shown on their twofold axes (lines).



Figure 3.

Structure of the porcine insulin monomer. The B‐chain peptide backbone is shown as a dark ribbon and the A chain as a light ribbon.



Figure 4.

A: Covalent structure of rat preproinsulin I.

B: Primary sequences of representative vertebrate proinsulins compared with the human sequence. Conserved residues are highlighted. Sources are as follows: human 17, hummingbird 68, Xenopus 265, zebraflsh 179, hagfish 39, amphioxus 38.



Figure 5.

A: Covalent structure of rat preproinsulin I.

B: Primary sequences of representative vertebrate proinsulins compared with the human sequence. Conserved residues are highlighted. Sources are as follows: human 17, hummingbird 68, Xenopus 265, zebraflsh 179, hagfish 39, amphioxus 38.



Figure 6.

Subcellular organization of the insulin biosynthetic pathway. Arrows indicate direction of movement of (pro)insulin via subcellular compartments. Dashed arrow pointing downward indicates the mannose‐6‐P receptor pathway to the lysosome. Dashed arrow pointing upward indicates granule degradative pathway (autophagy). Time scales indicate intracompartmental residence times (see text for details). RER, rough endoplasmic reticulum.



Figure 7.

Schematic diagram showing “constitutive‐like” alternative secretory pathway arising from immature secretory granules (vesicles) via passive sorting of soluble, nonaggregated C peptide generated by proinsulin processing during early phases of secretory granule maturation. Other granule constituents, such as furin and/or lysosomal enzyme precursors, may be actively removed from the maturing granules via clathrin‐clad vesicles by virtue of cytosolic domain interactions (furin) or mannose‐6‐P receptor (M‐6‐PR)–mediated pathways (cathepsins) (see text and ref. 6 for details). TGN, trans‐Golgi network.



Figure 8.

Proinsulin processing pathways in the β cell. On the right is the predominant pathway, beginning with cleavage of proinsulin by prohormone convertase 3 (PC3) to yield des‐31, 32‐proinsulin, a preferred substrate for PC2. However, evidence summarized in the text indicates that either enzyme is capable of cleavage at both junctions in the prohormone to generate insulin. Carboxy peptidase E (CPE) removes C‐terminal basic amino acids from products (not shown) generated by the endoproteolytic action of PC2 or PC3.



Figure 9.

Structures of the subtilisin‐like proprotein convertases. The upper three [Prohormone convertase 2 (PC2), PC1/PC3, and PC4] are neuroendocrine‐specific in their localization and functions, while the lower group (furin, PACE, PC6, and PC7) are expressed more widely and tend to process a variety of constitutively secreted precursors (for review, see ref. 234). Pre, signal peptide; Pro, pro domain; Cat, catalytic domain; P, P domain; CR cysteine‐rich domain; AH, amphipathic helix; TM, transmembrane segment; S/TR, serinethreonine‐rich region. Catalytic residues D, H, N, and S are shown above the catalytic domains. Glycosylation sites are not shown.



Figure 10.

Structure of the insulin gene in selected vertebrates, showing highly conserved intron–exon structure and the promoter region. Numbers indicate variable size (in nucleotides) of each intron.



Figure 11.

Electron micrograph of β cells in wild‐type (PC2 +/+) or mutant (PC‐/‐) mice. Note abundance of mature secretory granules in wild‐type contrasting with large numbers of immature‐appearing secretory granules in the mutant, indicative of increased proinsulin content 78. Bar = 1 mM.

(Photomicrograph courtesy of Hewson H. Swift.)
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Article Title: Biosynthesis of Insulin
 

How to Cite

Donald F. Steiner, Shu Jin Chan, Arthur H. Rubenstein. Biosynthesis of Insulin. Compr Physiol 2011, Supplement 21: Handbook of Physiology, The Endocrine System, The Endocrine Pancreas and Regulation of Metabolism: 49-78. First published in print 2001. doi: 10.1002/cphy.cp070203